Journal articles: 'Cell culture techniques'

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Morton, A. J. "Practical Cell Culture Techniques." Trends in Neurosciences 16, no. 6 (June 1993): 245–46. http://dx.doi.org/10.1016/0166-2236(93)90165-i.

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Ylostalo, Joni H. "3D Stem Cell Culture." Cells 9, no. 10 (September 27, 2020): 2178. http://dx.doi.org/10.3390/cells9102178.

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Much interest has been directed towards stem cells, both in basic and translational research, to understand basic stem cell biology and to develop new therapies for many disorders. In general, stem cells can be cultured with relative ease, however, most common culture methods for stem cells employ 2D techniques using plastic. These cultures do not well represent the stem cell niches in the body, which are delicate microenvironments composed of not only stem cells, but also supporting stromal cells, extracellular matrix, and growth factors. Therefore, researchers and clinicians have been seeking optimal stem cell preparations for basic research and clinical applications, and these might be attainable through 3D culture of stem cells. The 3D cultures recapitulate the in vivo cell-to-cell and cell-to-matrix interactions more effectively, and the cells in 3D cultures exhibit many unique and desirable characteristics. The culture of stem cells in 3D may employ various matrices or scaffolds, in addition to the cells, to support the complex structures. The goal of this Special Issue is to bring together recent research on 3D cultures of various stem cells to increase the basic understanding of stem cells and culture techniques, and also highlight stem cell preparations for possible novel therapeutic applications.

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Vààzquez, Joséé. "History of Cell Culture Techniques." American Biology Teacher 72, no. 8 (October 1, 2010): 518. http://dx.doi.org/10.1525/abt.2010.72.8.11.

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Shargool, Peter D. "Future uses of plant cell culture techniques." Biochemical Education 13, no. 2 (April 1985): 50–53. http://dx.doi.org/10.1016/0307-4412(85)90002-0.

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Greenberger, Joel S. "Combinatorial Cell Culture Techniques in Tissue Engineering." e-biomed: The Journal of Regenerative Medicine 1, no. 10 (October 24, 2000): 137–39. http://dx.doi.org/10.1089/152489000750009802.

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Phelan, Mary C. "Basic Techniques for Mammalian Cell Tissue Culture." Current Protocols in Cell Biology 00, no. 1 (October 1998): 1.1.1–1.1.10. http://dx.doi.org/10.1002/0471143030.cb0101s00.

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Marks, Edwin P., and Gordon B. Ward. "Cell culture techniques for studying insect cuticle." Archives of Insect Biochemistry and Physiology 6, no. 4 (December 1987): 217–25. http://dx.doi.org/10.1002/arch.940060403.

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Moran-Alvarez, Alba, Pedro Gonzalez-Menendez, Juan C. Mayo, and Rosa M. Sainz. "Reflections on the Biology of Cell Culture Models: Living on the Edge of Oxidative Metabolism in Cancer Cells." International Journal of Molecular Sciences 24, no. 3 (February 1, 2023): 2717. http://dx.doi.org/10.3390/ijms24032717.

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Nowadays, the study of cell metabolism is a hot topic in cancer research. Many studies have used 2D conventional cell cultures for their simplicity and the facility to infer mechanisms. However, the limitations of bidimensional cell cultures to recreate architecture, mechanics, and cell communication between tumor cells and their environment, have forced the development of other more realistic in vitro methodologies. Therefore, the explosion of 3D culture techniques and the necessity to reduce animal experimentation to a minimum has attracted the attention of researchers in the field of cancer metabolism. Here, we revise the limitations of actual culture models and discuss the utility of several 3D culture techniques to resolve those limitations.

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Bleotu, Coralia, Carmen Diaconu, Mihaela Chivu, Irina Alexiu, Simona Ruta, and Costin Cernescu. "Evaluation of TV cell line viral susceptibility using conventional cell culture techniques." Open Medicine 1, no. 1 (March 1, 2006): 12–22. http://dx.doi.org/10.2478/s11536-006-0007-x.

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AbstractDespite the fact that a lot of methods have been developed for rapid virus detection, classic cell culture is still “the golden standard”. The range of viruses that can be isolated and cultured in cell line systems is often limited by the susceptibility of cells to support viral replication. Since the primary cell culture, the best cellular system available to support replication of a large number of viruses, is very expensive and diffcult to obtain, cell lines, which are easier to manipulate, are commonly used for virus growth and isolation.In two previous papers we described the TV cell line initiated by our team from a laryngeal tumor, which harbors human papillomavirus (HPV) gene sequences. In this paper we analyze its capacity to support virus replication. Depending on the virus, different cytopathic effects were produced. Comparison of viral effect observed on this cell line with the effect obtained on other cell lines has been performed. This cell line might be used in the clinical virology laboratory for virus isolation.

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Smeda, Reid J., and Stephen C. Weller. "Plant Cell and Tissue Culture Techniques for Weed Science Research." Weed Science 39, no. 3 (September 1991): 497–504. http://dx.doi.org/10.1017/s0043174500073288.

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Tissue and cell culture offer weed scientists many opportunities to research herbicide effects on plants. This review will discuss examples in which plant cells grown in vitro have been used to study herbicide action. Plant cell and tissue culture have many advantages over the use of whole plants; however, several disadvantages that exist are discussed. Cell cultures can be established for most plant species and provide a relatively homogeneous system for studying herbicide action. Responses of plant cells to herbicides are usually correlated with responses at the whole plant level, and cells have the advantage of posing fewer physical barriers to herbicide uptake and translocation. Cell culture techniques discussed include: screening candidate herbicide compounds; investigating herbicide efficacy, mechanism of action, metabolism, and uptake; and ascertaining mechanisms of herbicide resistance, selecting for resistance, and regenerating crops.

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He, Lingjie, Cheng Zhao, Qi Xiao, Ju Zhao, Haifeng Liu, Jun Jiang, and Quanquan Cao. "Profiling the Physiological Roles in Fish Primary Cell Culture." Biology 12, no. 12 (November 21, 2023): 1454. http://dx.doi.org/10.3390/biology12121454.

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Fish primary cell culture has emerged as a valuable tool for investigating the physiological roles and responses of various cell types found in fish species. This review aims to provide an overview of the advancements and applications of fish primary cell culture techniques, focusing on the profiling of physiological roles exhibited by fish cells in vitro. Fish primary cell culture involves the isolation and cultivation of cells directly derived from fish tissues, maintaining their functional characteristics and enabling researchers to study their behavior and responses under controlled conditions. Over the years, significant progress has been made in optimizing the culture conditions, establishing standardized protocols, and improving the characterization techniques for fish primary cell cultures. The review highlights the diverse cell types that have been successfully cultured from different fish species, including gonad cells, pituitary cells, muscle cells, hepatocytes, kidney and immune cells, adipocyte cells and myeloid cells, brain cells, primary fin cells, gill cells, and other cells. Each cell type exhibits distinct physiological functions, contributing to vital processes such as metabolism, tissue regeneration, immune response, and toxin metabolism. Furthermore, this paper explores the pivotal role of fish primary cell culture in elucidating the mechanisms underlying various physiological processes. Researchers have utilized fish primary cell cultures to study the effects of environmental factors, toxins, pathogens, and pharmaceutical compounds on cellular functions, providing valuable insights into fish health, disease pathogenesis, and drug development. The paper also discusses the application of fish primary cell cultures in aquaculture research, particularly in investigating fish growth, nutrition, reproduction, and stress responses. By mimicking the in vivo conditions in vitro, primary cell culture has proven instrumental in identifying key factors influencing fish health and performance, thereby contributing to the development of sustainable aquaculture practices.

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Lovitt, Carrie, Todd Shelper, and Vicky Avery. "Advanced Cell Culture Techniques for Cancer Drug Discovery." Biology 3, no. 2 (May 30, 2014): 345–67. http://dx.doi.org/10.3390/biology3020345.

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Smith-Thomas, L. C., and J. W. Fawcett. "Expression of Schwann cell markers by mammalian neural crest cells in vitro." Development 105, no. 2 (February 1, 1989): 251–62. http://dx.doi.org/10.1242/dev.105.2.251.

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During embryonic development, neural crest cells differentiate into a wide variety of cell types including Schwann cells of the peripheral nervous system. In order to establish when neural crest cells first start to express a Schwann cell phenotype immunocytochemical techniques were used to examine rat premigratory neural crest cell cultures for the presence of Schwann cell markers. Cultures were fixed for immunocytochemistry after culture periods ranging from 1 to 24 days. Neural crest cells were identified by their morphology and any neural tube cells remaining in the cultures were identified by their epithelial morphology and immunocytochemically. As early as 1 to 2 days in culture, approximately one third of the neural crest cells stained with m217c, a monoclonal antibody that appears to recognize the same antigen as rat neural antigen-1 (RAN-1). A similar proportion of cells were immunoreactive in cultures stained with 192-IgG, a monoclonal antibody that recognizes the rat nerve growth factor receptor. The number of immunoreactive cells increased with time in culture. After 16 days in culture, nests of cells, many of which had a bipolar morphology, were present in the area previously occupied by neural crest cells. The cells in the nests were often associated with neurons and were immunoreactive for m217c, 192-IgG and antibody to S-100 protein and laminin, indicating that the cells were Schwann cells. At all culture periods examined, neural crest cells did not express glial fibrillary acidic protein. These results demonstrate that cultured premigratory neural crest cells express early Schwann cell markers and that some of these cells differentiate into Schwann cells. These observations suggest that some neural crest cells in vivo may be committed to forming Schwann cells and will do so provided that they then proceed to encounter the correct environmental cues during embryonic development.

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Zuščíková, Lucia, Hana Greifová, Denis Bažány, Norbert Lukáč, and Tomáš Jambor. "Current approaches and techniques of 3D cell culture systems: a review." Archives of Ecotoxicology 6, no. 1 (June 29, 2024): 22–27. http://dx.doi.org/10.36547/ae.2024.6.1.22-27.

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Presently, the majority of cells are cultivated by two-dimensional (2D) methods; however, latest and enhanced procedures employing three-dimensional (3D) cell culturing techniques provide strong indications that significantly more sophisticated studies can be carried out, providing invaluable insights. Recent years have seen a rapid development of 3D cell cultures since cells grown in a static environment on a flat substrate are far from reaching an in vivo condition. Currently, scientists are gradually realising that in vitro cell shape, structure, and physiological activities may be achieved. As a resolution, a three-dimensional matrix-like framework for cell attachment, proliferation, differentiation, and communication in both static and dynamic culture conditions is what three-dimensional cell carriers have gradually come to offer. Different mechanical stimulations that more closely resemble the genuine in vivo microenvironment could be the main function of 3D cell carriers in dynamic culture systems. Current developments in 3D dynamic cell culture techniques have been presented in this review, along with a discussion of their benefits and drawbacks when compared to conventional 2D cell growth under static settings.

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Vermorken, A. J. M. "Culture Techniques and Potential Research Applications of Human Hair Follicle Cells In Vitro." Alternatives to Laboratory Animals 13, no. 1 (September 1985): 8–37. http://dx.doi.org/10.1177/026119298501300104.

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Since 1980, when human hair follicle cells were cultured in vitro for the first time, a whole series of techniques have been developed that render hair follicle keratinocytes as easy to handle in culture as fibroblasts. As a consequence, one can conclude that the need for a method providing for the routine cultivation of easily obtainable human primary epithelial cells has now been met, and it may be expected that more and more workers will use hair follicle keratinocytes for studies that specifically require human epithelial cells. The ease of culture and the ready availability of material may encourage workers to consider human hair follicle cell culture before either animal models or cultures of cells derived from invasive skin biopsies.

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Ceresa, Claudia C., Alan J. Knox, and Simon R. Johnson. "Use of a three-dimensional cell culture model to study airway smooth muscle-mast cell interactions in airway remodeling." American Journal of Physiology-Lung Cellular and Molecular Physiology 296, no. 6 (June 2009): L1059—L1066. http://dx.doi.org/10.1152/ajplung.90445.2008.

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Increased airway smooth muscle (ASM) mass and infiltration by mast cells are key features of airway remodeling in asthma. We describe a model to investigate the relationship between ASM, the extracellular matrix, mast cells, and airway remodeling. ASM cells were cultured in a three-dimensional (3-D) collagen I gel (3-D culture) alone or with mast cells. Immunocytochemistry and Western blotting of ASM in 3-D cultures revealed a spindle-shaped morphology and significantly lower α-smooth muscle actin and vimentin expression than in ASM cultured in monolayers on collagen type I or plastic (2-D culture). In 3-D cultures, basal ASM proliferation, examined by Ki67 immunocytochemistry, was reduced to 33 ± 7% ( P < 0.05) of that in 2-D cultures. The presence of mast cells in cocultures increased ASM proliferation by 1.8-fold ( P < 0.05). Gelatin zymography revealed more active matrix metalloproteinase (MMP)-2 in 3-D than in 2-D culture supernatants over 7 days. Functional MMP activity was examined by gel contraction. The spontaneous gel contraction over 7 days was significantly inhibited by the MMP inhibitor ilomastat. Mast cell coculture enhanced ASM gel contraction by 22 ± 16% (not significant). Our model shows that ASM has different morphology, with lower contractile protein expression and basal proliferation in 3-D culture. Compared with standard techniques, ASM synthetic function, as shown by MMP production and activity, is sustained over longer periods. The presence of mast cells in the 3-D model enhanced ASM proliferation and MMP production. Airway remodeling in asthma may be more accurately modeled by our system than by standard culture systems.

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Drigas, Athanasios, and Maria Pouliou. "E-Culture Techniques and Applications." International Journal of Knowledge Society Research 4, no. 4 (October 2013): 11–17. http://dx.doi.org/10.4018/ijksr.2013100102.

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E-culture is the combination of Information and Communication Technologies (ICTs) with traditional culture. Cultural heritage reveals elements of the past, which renders the occupation with it by contemporary societies, an absolute necessity. This paper aims to present a thorough review of e-culture, its methods and its applications in recent years. First, it focuses on the alternative choice of creating virtual museums in order to improve traditional museums' services through an attractive way to the visitor. Moreover, it discusses the significance of cultural digitization analysing the methods of digitization for both monuments and objects. Finally, it mentions the potential of numerous existing guide applications, which can be installed on cell phones and whose aim is to facilitate the navigation of visitors at archaeological sites and museums, as well as the potential of robots, which guide visitors to museums in an interactive way.

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Wu, Ying-ying, Yu Ban, Ning Geng, Yong-yue Wang, Xiao-guang Liu, Tao Yu, and Ping Gong. "Evaluation of different culture techniques of osteoblasts on 3D scaffolds." Open Life Sciences 5, no. 4 (August 1, 2010): 456–65. http://dx.doi.org/10.2478/s11535-010-0027-z.

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AbstractBones adjust their structure to withstand the mechanical demands they experience. It is suggested that flow-derived shear stress may be the most significant and primary mediator of mechanical stimulation. In this study, we designed and fabricated a fluid flow cell culture system that can load shear stress onto cells cultured on 3D scaffolds. We evaluated the effect of different culture techniques, namely, (1) continuous perfusion fluid flow, (2) intermittent perfusion fluid flow, and (3) static condition, on the proliferation of osteoblasts seeded on partially deproteinized bones. The flow rate was set at 1 ml/min for all the cells cultured using flow perfusion and the experiment was conducted for 12 days. Scanning electron microscopy analysis indicated an increase in cell proliferation for scaffolds subjected to fluid shear stress. In addition, the long axes of these cells lengthened along the flowing fluid direction. Continuous perfusion significantly enhanced cell proliferation compared to either intermittent perfusion or static condition. All the results demonstrated that fluid shear stress is able to enhance the proliferation of cells and change the form of cells.

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Shyam, Rohin, L. Reddy, and Arunkumar Palaniappan. "Fabrication and Characterization Techniques of In Vitro 3D Tissue Models." International Journal of Molecular Sciences 24, no. 3 (January 18, 2023): 1912. http://dx.doi.org/10.3390/ijms24031912.

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The culturing of cells in the laboratory under controlled conditions has always been crucial for the advancement of scientific research. Cell-based assays have played an important role in providing simple, fast, accurate, and cost-effective methods in drug discovery, disease modeling, and tissue engineering while mitigating reliance on cost-intensive and ethically challenging animal studies. The techniques involved in culturing cells are critical as results are based on cellular response to drugs, cellular cues, external stimuli, and human physiology. In order to establish in vitro cultures, cells are either isolated from normal or diseased tissue and allowed to grow in two or three dimensions. Two-dimensional (2D) cell culture methods involve the proliferation of cells on flat rigid surfaces resulting in a monolayer culture, while in three-dimensional (3D) cell cultures, the additional dimension provides a more accurate representation of the tissue milieu. In this review, we discuss the various methods involved in the development of 3D cell culture systems emphasizing the differences between 2D and 3D systems and methods involved in the recapitulation of the organ-specific 3D microenvironment. In addition, we discuss the latest developments in 3D tissue model fabrication techniques, microfluidics-based organ-on-a-chip, and imaging as a characterization technique for 3D tissue models.

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Tekkatte, Chandana, Gency Ponrose Gunasingh, K. M. Cherian, and Kavitha Sankaranarayanan. "“Humanized” Stem Cell Culture Techniques: The Animal Serum Controversy." Stem Cells International 2011 (2011): 1–14. http://dx.doi.org/10.4061/2011/504723.

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Cellular therapy is reaching a pinnacle with an understanding of the potential of human mesenchymal stem cells (hMSCs) to regenerate damaged tissue in the body. The limited numbers of these hMSCs in currently identified sources, like bone marrow, adipose tissue, and so forth, bring forth the need for theirin vitroculture/expansion. However, the extensive usage of supplements containing xenogeneic components in the expansion-media might pose a risk to the post-transplantation safety of patients. This warrants the necessity to identify and develop chemically defined or “humanized” supplements which would makein vitrocultured/processed cells relatively safer for transplantation in regenerative medicine. In this paper, we outline the various caveats associated with conventionally used supplements of xenogenic origin and also portray the possible alternatives/additives which could one day herald the dawn of a new era in the translation ofin vitrocultured cells to therapeutic interventions.

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Wilson, G. "Cell culture techniques for the study of drug transport." European Journal of Drug Metabolism and Pharmacokinetics 15, no. 2 (April 1990): 159–63. http://dx.doi.org/10.1007/bf03190199.

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Mozdziak, Paul E., James N. Petitte, and Susan D. Carson. "An introductory undergraduate course covering animal cell culture techniques." Biochemistry and Molecular Biology Education 32, no. 5 (September 2004): 319–22. http://dx.doi.org/10.1002/bmb.2004.494032050381.

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Barozzi, Dafne, and Cristina Scielzo. "Emerging Strategies in 3D Culture Models for Hematological Cancers." HemaSphere 7, no. 8 (July 27, 2023): e932. http://dx.doi.org/10.1097/hs9.0000000000000932.

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In vitro cell cultures are fundamental and necessary tools in cancer research and personalized drug discovery. Currently, most cells are cultured using two-dimensional (2D) methods, and drug testing is mainly performed in animal models. However, new and improved methods that implement three-dimensional (3D) cell-culturing techniques provide compelling evidence that more advanced experiments can be performed, yielding valuable new insights. In 3D cell-culture experiments, the cell environment can be manipulated to mimic the complexity and dynamicity of the human tissue microenvironment, possibly leading to more accurate representations of cell-to-cell interactions, tumor biology, and predictions of drug response. The 3D cell cultures can also potentially provide alternative ways to study hematological cancers and are expected to eventually bridge the gap between 2D cell culture and animal models. The present review provides an overview of the complexity of the lymphoid microenvironment and a summary of the currently used 3D models that aim at recreating it for hematological cancer research. We here dissect the differences and challenges between, and potential advantages of, different culture methods and present our vision of the most promising future strategies in the hematological field.

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Amini, M., J. Hisdal, and H. Kalvøy. "Applications of bioimpedance measurement techniques in tissue engineering." Journal of Electrical Bioimpedance 9, no. 1 (December 31, 2018): 142–58. http://dx.doi.org/10.2478/joeb-2018-0019.

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Abstract Rapid development in the field of tissue engineering necessitates implementation of monitoring methods for evaluation of the viability and characteristics of the cell cultures in a real-time, non-invasive and non-destructive manner. Current monitoring techniques are mainly histological and require labeling and involve destructive tests to characterize cell cultures. Bioimpedance measurement technique which benefits from measurement of electrical properties of the biological tissues, offers a non-invasive, label-free and real-time solution for monitoring tissue engineered constructs. This review outlines the fundamentals of bioimpedance, as well as electrical properties of the biological tissues, different types of cell culture constructs and possible electrode configuration set ups for performing bioimpedance measurements on these cell cultures. In addition, various bioimpedance measurement techniques and their applications in the field of tissue engineering are discussed.

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Gunti, Sreenivasulu, Austin T. K. Hoke, Kenny P. Vu, and Nyall R. London. "Organoid and Spheroid Tumor Models: Techniques and Applications." Cancers 13, no. 4 (February 19, 2021): 874. http://dx.doi.org/10.3390/cancers13040874.

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Techniques to develop three-dimensional cell culture models are rapidly expanding to bridge the gap between conventional cell culture and animal models. Organoid and spheroid cultures have distinct and overlapping purposes and differ in cellular sources and protocol for establishment. Spheroids are of lower complexity structurally but are simple and popular models for drug screening. Organoids histologically and genetically resemble the original tumor from which they were derived. Ease of generation, ability for long-term culture and cryopreservation make organoids suitable for a wide range of applications. Organoids-on-chip models combine organoid methods with powerful designing and fabrication of micro-chip technology. Organoid-chip models can emulate the dynamic microenvironment of tumor pathophysiology as well as tissue–tissue interactions. In this review, we outline different tumor spheroid and organoid models and techniques to establish them. We also discuss the recent advances and applications of tumor organoids with an emphasis on tumor modeling, drug screening, personalized medicine and immunotherapy.

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Jarrahy, Reza, Weibiao Huang, George H. Rudkin, Jane M. Lee, Kenji Ishida, Micah D. Berry, Modar Sukkarieh, Benjamin M. Wu, Dean T. Yamaguchi, and Timothy A. Miller. "Osteogenic differentiation is inhibited and angiogenic expression is enhanced in MC3T3-E1 cells cultured on three-dimensional scaffolds." American Journal of Physiology-Cell Physiology 289, no. 2 (August 2005): C408—C414. http://dx.doi.org/10.1152/ajpcell.00196.2004.

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Osteogenic differentiation of osteoprogenitor cells in three-dimensional (3D) in vitro culture remains poorly understood. Using quantitative real-time RT-PCR techniques, we examined mRNA expression of alkaline phosphatase, osteocalcin, and vascular endothelial growth factor (VEGF) in murine preosteoblastic MC3T3-E1 cells cultured for 48 h and 14 days on conventional two-dimensional (2D) poly(l-lactide-co-glycolide) (PLGA) films and 3D PLGA scaffolds. Differences in VEGF secretion and function between 2D and 3D culture systems were examined using Western blots and an in vitro Matrigel-based angiogenesis assay. Expression of both alkaline phosphatase and osteocalcin in cells cultured on 3D scaffolds was significantly downregulated relative to 2D controls in 48 h and 14 day cultures. In contrast, elevated levels of VEGF expression in 3D culture were noted at every time point in short- and long-term culture. VEGF protein secretion in 3D cultures was triple the amount of secretion observed in 2D controls. Conditioned medium from 3D cultures induced an enhanced level of angiogenic activity, as evidenced by increases in branch points observed in in vitro angiogenesis assays. These results collectively indicate that MC3T3-E1 cells commit to osteogenic differentiation at a slower rate when cultured on 3D PLGA scaffolds and that VEGF is preferentially expressed by these cells when they are cultured in three dimensions.

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Rai, Abdelwahab, Zohra Ammi, Dahbia Leila Anes-Boulahbal, Aymen Amin Assadi, Abdeltif Amrane, Oussama Baaloudj, and Lotfi Mouni. "Molecular Amplification and Cell Culturing Efficiency for Enteroviruses’ Detection in Cerebrospinal Fluids of Algerian Patients Suffering from Meningitis." Viruses 16, no. 2 (January 23, 2024): 170. http://dx.doi.org/10.3390/v16020170.

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Enteroviruses (EVs) represent a major cause of viral meningitis, being responsible for nearly 1 billion infections each year worldwide. Several techniques were developed to obtain better diagnostic results of EV infections. Herein, we evaluated the efficiency of EV detection through isolation on both Rhabdomyosarcoma (RD) and Vero cell line cultures, conventional reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR. Thus, 50 cerebrospinal fluid (CSF) samples belonging to patients suspected to have viral meningitis in northern Algeria were collected, anonymously numbered from 1 to 50 and subjected to the above-mentioned techniques for EV detection. Using real-time RT-PCR, 34 CSF samples were revealed to be positive for viral origin of meningitis (68%). Thirteen of them were positive when the conventional RT-PCR was used (26%), and only three samples gave positive results when the cell culture technique was used (6%). Surprisingly, two cell culture-positive CSF samples, namely, 31 and 39, were negative using RT-PCR directly on the original samples. However, they turned to be positive when amplification was carried out on their corresponding cell culture supernatant. The cell-cultured viral isolates were then identified by sequencing their viral genome’s VP1 regions. All of them were revealed to belong to the echovirus 27 strain. This investigation demonstrates that RT-PCR techniques are often more sensitive, accurate and much faster, providing reliable results within a clinically acceptable timeframe. However, viral isolation on cell cultures remains crucial to obtain enough viral load for serological tests or even to avoid the rare, but existing, false negative PCR.

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Chitturi Suryaprakash, Ravi Teja, Omar Kujan, Kate Shearston, and Camile S. Farah. "Three-Dimensional Cell Culture Models to Investigate Oral Carcinogenesis: A Scoping Review." International Journal of Molecular Sciences 21, no. 24 (December 14, 2020): 9520. http://dx.doi.org/10.3390/ijms21249520.

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Three-dimensional (3-D) cell culture models, such as spheroids, organoids, and organotypic cultures, are more physiologically representative of the human tumor microenvironment (TME) than traditional two-dimensional (2-D) cell culture models. They have been used as in vitro models to investigate various aspects of oral cancer but, to date, have not be widely used in investigations of the process of oral carcinogenesis. The aim of this scoping review was to evaluate the use of 3-D cell cultures in oral squamous cell carcinoma (OSCC) research, with a particular emphasis on oral carcinogenesis studies. Databases (PubMed, Scopus, and Web of Science) were systematically searched to identify research applying 3-D cell culture techniques to cells from normal, dysplastic, and malignant oral mucosae. A total of 119 studies were included for qualitative analysis including 53 studies utilizing spheroids, 62 utilizing organotypic cultures, and 4 using organoids. We found that 3-D oral carcinogenesis studies had been limited to just two organotypic culture models and that to date, spheroids and organoids had not been utilized for this purpose. Spheroid culture was most frequently used as a tumorosphere forming assay and the organoids cultured from human OSCCs most often used in drug sensitivity testing. These results indicate that there are significant opportunities to utilize 3-D cell culture to explore the development of oral cancer, particularly as the physiological relevance of these models continues to improve.

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Wongvisavavit, Rintra, Mohit Parekh, Sajjad Ahmad, and Julie T. Daniels. "Challenges in corneal endothelial cell culture." Regenerative Medicine 16, no. 9 (September 2021): 871–91. http://dx.doi.org/10.2217/rme-2020-0202.

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Corneal endothelial cells (CECs) facilitate the function of maintaining the transparency of the cornea. Damage or dysfunction of CECs can lead to blindness, and the primary treatment is corneal transplantation. However, the shortage of cornea donors is a significant problem worldwide. Thus, cultured CEC therapy has been proposed and found to be a promising approach to overcome the lack of tissue supply. Unfortunately, CECs in humans rarely proliferate in vivo and, therefore, can be extremely challenging to culture in vitro. Several promising cell isolation and culture techniques have been proposed. Multiple factors affecting the success of cell expansion including donor characteristics, preservation and isolation methods, plating density, media preparation, transdifferentiation and biomarkers have been evaluated. However, there is no consensus on standard technique for CEC culture. This review aimed to determine the challenges and investigate potential options that would facilitate the standardization of CEC culture for research and therapeutic application.

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Badr-Eldin, Shaimaa M., Hibah M. Aldawsari, Sabna Kotta, Pran Kishore Deb, and Katharigatta N. Venugopala. "Three-Dimensional In Vitro Cell Culture Models for Efficient Drug Discovery: Progress So Far and Future Prospects." Pharmaceuticals 15, no. 8 (July 27, 2022): 926. http://dx.doi.org/10.3390/ph15080926.

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Despite tremendous advancements in technologies and resources, drug discovery still remains a tedious and expensive process. Though most cells are cultured using 2D monolayer cultures, due to lack of specificity, biochemical incompatibility, and cell-to-cell/matrix communications, they often lag behind in the race of modern drug discovery. There exists compelling evidence that 3D cell culture models are quite promising and advantageous in mimicking in vivo conditions. It is anticipated that these 3D cell culture methods will bridge the translation of data from 2D cell culture to animal models. Although 3D technologies have been adopted widely these days, they still have certain challenges associated with them, such as the maintenance of a micro-tissue environment similar to in vivo models and a lack of reproducibility. However, newer 3D cell culture models are able to bypass these issues to a maximum extent. This review summarizes the basic principles of 3D cell culture approaches and emphasizes different 3D techniques such as hydrogels, spheroids, microfluidic devices, organoids, and 3D bioprinting methods. Besides the progress made so far in 3D cell culture systems, the article emphasizes the various challenges associated with these models and their potential role in drug repositioning, including perspectives from the COVID-19 pandemic.

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Sharti, Mojtaba, Hadi E. G. Ghaleh, Ruhollah Dorostkar, Mehdi Tat, Ali Razei, Alireza Rafati, and Akbar G. Alvanegh. "Introducing two effective techniques for the detection of adenovirus respiratory infection in human sample." Romanian Journal of Military Medicine 124, no. 4 (January 11, 2021): 488–92. http://dx.doi.org/10.55453/rjmm.2021.124.4.11.

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Recently, respiratory viral infections have witnessed substantial diagnostic evolution, with the advent of emergent pathogenic agents and the improvement of new diagnostic approaches. Acute viral respiratory infection (ARVI) commonly causes illness and fatality, particularly in pediatric patients. Adenovirus has the highest prevalence in upper and lower respiratory tract infections (RTIs) amongst respiratory pathogens of importance. The purpose of this research was to introduce two effective techniques for the detection of adenovirus respiratory infection in Human samples, cell culture, and real-time PCR. Samples from patients aged less than ten years were obtained from Baqiyatallah (as) Hospital and cultured on HEK cells after preparation. After observing the cytopathic effects of the virus, a molecular test (Real-time PCR) with the help of specific primers was done on samples taken from the cell culture. Both assays detected viruses in clinical samples, and results of cell culture and real-time PCR confirmed infection with adenovirus in samples. According to our findings, simultaneous use of tests (cell culture and PCR) based test real-time PCR is a somewhat reliable method for detecting adenovirus in clinical specimens.

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Du, Eric Y., Adam D. Martin, Celine Heu, and Pall Thordarson. "The Use of Hydrogels as Biomimetic Materials for 3D Cell Cultures." Australian Journal of Chemistry 70, no. 1 (2017): 1. http://dx.doi.org/10.1071/ch16241.

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With the recent developments in cell cultures and biomimetic materials, there is growing evidence indicating that long-established two-dimensional (2D) cell culture techniques are slowly being phased out and replaced with three-dimensional (3D) cell cultures. This is due to the 3D cell cultures better mimicking the natural extracellular matrix (ECM) where cells are found. The emergence of self-assembled hydrogels as an ECM mimic has revolutionised the field owing to their ability to closely simulate the fibrous nature of the ECM. Here, we review recent progress in using hydrogels as biomimetic materials in 3D cell cultures, particularly supramolecular peptide hydrogels. With greater comprehension of the behaviour of cells in these hydrogels, a cell culture system that can be used in a wide array of 3D culture-based applications can be developed.

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Kamp-Glass, Marihelen. "TEACHING PLANT TISSUE CULTURE TECHNIQUES AND EXPERIMENTS." HortScience 28, no. 5 (May 1993): 495c—495. http://dx.doi.org/10.21273/hortsci.28.5.495c.

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With the expansion of agriculture biotechnology, plant biotechnology has become increasingly popular as a means of producing commercially useful crops for consumption and research. The ability to perform research on crops requires a basic understanding of the techniques of plant cell and tissue culture for genetics and other areas of plant physiology and horticulture. Teaching basic plant tissue culture techniques such as aseptic techniques, media preparation, explant orientation, callus induction and embryo culture will be discussed.

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Leland, Diane S., and Christine C. Ginocchio. "Role of Cell Culture for Virus Detection in the Age of Technology." Clinical Microbiology Reviews 20, no. 1 (January 2007): 49–78. http://dx.doi.org/10.1128/cmr.00002-06.

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SUMMARY Viral disease diagnosis has traditionally relied on the isolation of viral pathogens in cell cultures. Although this approach is often slow and requires considerable technical expertise, it has been regarded for decades as the “gold standard” for the laboratory diagnosis of viral disease. With the development of nonculture methods for the rapid detection of viral antigens and/or nucleic acids, the usefulness of viral culture has been questioned. This review describes advances in cell culture-based viral diagnostic products and techniques, including the use of newer cell culture formats, cryopreserved cell cultures, centrifugation-enhanced inoculation, precytopathogenic effect detection, cocultivated cell cultures, and transgenic cell lines. All of these contribute to more efficient and less technically demanding viral detection in cell culture. Although most laboratories combine various culture and nonculture approaches to optimize viral disease diagnosis, virus isolation in cell culture remains a useful approach, especially when a viable isolate is needed, if viable and nonviable virus must be differentiated, when infection is not characteristic of any single virus (i.e., when testing for only one virus is not sufficient), and when available culture-based methods can provide a result in a more timely fashion than molecular methods.

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Craig, B., L. Hawkey, and A. LeFurgey. "Techniques for cryoultramicrotomy of propane jet frozen biological samples." Proceedings, annual meeting, Electron Microscopy Society of America 44 (August 1986): 260–61. http://dx.doi.org/10.1017/s042482010014292x.

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Ultra-rapid freezing followed by cryoultramicrotomy is essential for the preservation of diffusible elements in situ within cells prior to scanning transmission electron microscopy and quantitative energy dispersive x-ray microanalysis. For cells or tissue fragments in suspension and for monolayer cell cultures, propane jet freezing provides cooling rates greater than 30,000°C/sec with regions up to 40μm in thickness free of significant ice crystal formation. While this method of freezing has frequently been applied prior to freeze fracture or freeze substitution, it has not been widely utilized prior to cryoultramicrotomy and subsequent x-ray microanalytical studies. This report describes methods devised in our laboratory for cryosectioning of propane jet frozen kidney proximal tubule suspensions and cultured embryonic chick heart cells, in particular a new technique for mounting frozen suspension specimens for sectioning. The techniques utilize the same specimen supports and sample holders as those used for freeze fracture and freeze substitution and should be generally applicable to any cell suspension or culture preparation.

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Kammerer, Sarah. "Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro." International Journal of Molecular Sciences 22, no. 19 (September 23, 2021): 10214. http://dx.doi.org/10.3390/ijms221910214.

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Drug-induced liver injury (DILI) is the major reason for failures in drug development and withdrawal of approved drugs from the market. Two-dimensional cultures of hepatocytes often fail to reliably predict DILI: hepatoma cell lines such as HepG2 do not reflect important primary-like hepatic properties and primary human hepatocytes (pHHs) dedifferentiate quickly in vitro and are, therefore, not suitable for long-term toxicity studies. More predictive liver in vitro models are urgently required in drug development and compound safety evaluation. This review discusses available human hepatic cell types for in vitro toxicology analysis and their usage in established and emerging three-dimensional (3D) culture systems. Generally, 3D cultures maintain or improve primary hepatic functions (including expression of drug-metabolizing enzymes) of different liver cells for several weeks of culture, thus allowing long-term and repeated-dose toxicity studies. Spheroid cultures of pHHs have been comprehensively tested, but also other cell types such as HepaRG benefit from 3D culture systems. Emerging 3D culture techniques include usage of induced pluripotent stem-cell-derived hepatocytes and primary-like upcyte cells, as well as advanced culture techniques such as microfluidic liver-on-a-chip models. In-depth characterization of existing and emerging 3D hepatocyte technologies is indispensable for successful implementation of such systems in toxicological analysis.

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Valentich, J. D., and M. F. Stokols. "An established cell line from mouse kidney medullary thick ascending limb. I. Cell culture techniques, morphology, and antigenic expression." American Journal of Physiology-Cell Physiology 251, no. 2 (August 1, 1986): C299—C311. http://dx.doi.org/10.1152/ajpcell.1986.251.2.c299.

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The effective use of cultured cells as model systems for investigating the differentiation and regulation of transport processes in renal tubular epithelial cells depends on the availability of functional long-term cell lines derived from specific nephron segments. Conventional culture procedures that treat cells as proliferating microorganisms possess several inherent limitations that could contribute to phenotypic instability and limited proliferative capacity in vitro. In this study, culture techniques were adopted that avoid exposure of cells to proteolytic enzymes, maintain intercellular contacts, and allow cells to remain continually adherent to a collagen gel substratum. This methodology resulted in the development of a continuous epithelial cell line from identified, microdissected segments of the mouse kidney medullary thick ascending limb.

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van Rijt, Antonia, Evan Stefanek, and Karolina Valente. "Preclinical Testing Techniques: Paving the Way for New Oncology Screening Approaches." Cancers 15, no. 18 (September 7, 2023): 4466. http://dx.doi.org/10.3390/cancers15184466.

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Prior to clinical trials, preclinical testing of oncology drug candidates is performed by evaluating drug candidates with in vitro and in vivo platforms. For in vivo testing, animal models are used to evaluate the toxicity and efficacy of drug candidates. However, animal models often display poor translational results as many drugs that pass preclinical testing fail when tested with humans, with oncology drugs exhibiting especially poor acceptance rates. The FDA Modernization Act 2.0 promotes alternative preclinical testing techniques, presenting the opportunity to use higher complexity in vitro models as an alternative to in vivo testing, including three-dimensional (3D) cell culture models. Three-dimensional tissue cultures address many of the shortcomings of 2D cultures by more closely replicating the tumour microenvironment through a combination of physiologically relevant drug diffusion, paracrine signalling, cellular phenotype, and vascularization that can better mimic native human tissue. This review will discuss the common forms of 3D cell culture, including cell spheroids, organoids, organs-on-a-chip, and 3D bioprinted tissues. Their advantages and limitations will be presented, aiming to discuss the use of these 3D models to accurately represent human tissue and as an alternative to animal testing. The use of 3D culture platforms for preclinical drug development is expected to accelerate as these platforms continue to improve in complexity, reliability, and translational predictivity.

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Srikanth, ChitturV, Gaur Preksha, and Rajendran Yesheswini. "Cell culture techniques in gastrointestinal research: Methods, possibilities and challenges." Indian Journal of Pathology and Microbiology 64, no. 5 (2021): 52. http://dx.doi.org/10.4103/ijpm.ijpm_933_20.

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Di Iorio, Enzo, Stefano Ferrari, Adriano Fasolo, Elisabetta Böhm, Diego Ponzin, and Vanessa Barbaro. "Techniques for Culture and Assessment of Limbal stem Cell Grafts." Ocular Surface 8, no. 3 (July 2010): 146–53. http://dx.doi.org/10.1016/s1542-0124(12)70225-2.

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Dalton, Colin C. "Handbook of plant cell culture vol. 4 techniques and applications." Trends in Genetics 3 (January 1987): 172. http://dx.doi.org/10.1016/0168-9525(87)90220-4.

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Lock, S. O., and J. V. Friend. "Phototoxicity testing In vitro: Evaluation of mammalian cell culture techniques." Food and Chemical Toxicology 24, no. 6-7 (June 1986): 789–93. http://dx.doi.org/10.1016/0278-6915(86)90187-0.

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Bhojwani, Sant S. "Handbook of plant cell culture, vol. 4. Techniques and applications." Scientia Horticulturae 44, no. 3-4 (November 1990): 347–48. http://dx.doi.org/10.1016/0304-4238(90)90136-3.

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Tan, Lu, and Kristin Schirmer. "Cell culture-based biosensing techniques for detecting toxicity in water." Current Opinion in Biotechnology 45 (June 2017): 59–68. http://dx.doi.org/10.1016/j.copbio.2016.11.026.

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Doherty, P. J. "Biocompatibility evaluation of glass ionomer cement using cell culture techniques." Clinical Materials 7, no. 4 (January 1991): 335–40. http://dx.doi.org/10.1016/0267-6605(91)90078-t.

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Shrestha, Sunil, Vinod Kumar Reddy Lekkala, Prabha Acharya, Darshita Siddhpura, and Moo-Yeal Lee. "Recent advances in microarray 3D bioprinting for high-throughput spheroid and tissue culture and analysis." Essays in Biochemistry 65, no. 3 (August 2021): 481–89. http://dx.doi.org/10.1042/ebc20200150.

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Abstract Three-dimensional (3D) cell culture in vitro has proven to be more physiologically relevant than two-dimensional (2D) culture of cell monolayers, thus more predictive in assessing efficacy and toxicity of compounds. There have been several 3D cell culture techniques developed, which include spheroid and multicellular tissue cultures. Cell spheroids have been generated from single or multiple cell types cultured in ultralow attachment (ULA) well plates and hanging droplet plates. In general, cell spheroids are formed in a relatively short period of culture, in the absence of extracellular matrices (ECMs), via gravity-driven self-aggregation, thus having limited ability to self-organization in layered structure. On the other hand, multicellular tissue cultures including miniature tissues derived from pluripotent stem cells and adult stem cells (a.k.a. ‘organoids’) and 3D bioprinted tissue constructs require biomimetic hydrogels or ECMs and show highly ordered structure due to spontaneous self-organization of cells during differentiation and maturation processes. In this short review article, we summarize traditional methods of spheroid and multicellular tissue cultures as well as their technical challenges, and introduce how droplet-based, miniature 3D bioprinting (‘microarray 3D bioprinting’) can be used to improve assay throughput and reproducibility for high-throughput, predictive screening of compounds. Several platforms including a micropillar chip and a 384-pillar plate developed to facilitate miniature spheroid and tissue cultures via microarray 3D bioprinting are introduced. We excluded microphysiological systems (MPSs) in this article although they are important tissue models to simulate multiorgan interactions.

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Rahman, MA, and MA Bari. "Callus induction and cell culture of castor (Ricinus communis L. CV. Shabje)." Journal of Bio-Science 20 (January 19, 2014): 161–69. http://dx.doi.org/10.3329/jbs.v20i0.17738.

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Context: Tissue culture is an important tool in plant biotechnology that allows for an increase in biomass or metabolite production by utilizing several techniques in callus or cell cultures. Ricin is a toxic protein that can be extracted from the castor bean as secondary metabolite. The ricin has been used experimentally in medicine to kill cancer cell. We developed callus and cell culture technique for the possible extraction of ricin from the cell suspension culture of Ricinus communis. Objectives: The present investigation aimed to develop the cell culture technique of R. communis L. cv. Shabje and tried to establish a protocol for cell suspension culture of castor for possible extraction of ricin from cell extract. Materials and Methods: The hypocotyl explants of R. communis L. cv. Shabje were used as experimental materials. Cultured on Murashige and Skoog medium supplemented with different concentrations and combinations of BAP, NAA, 2,4-D and IAA for callus induction. For cell culture, the media were used without agar with different concentrations and combinations of these hormones. Results: For callus induction the combination of BAP 2.0 mg/l + 0.5 mg/l NAA showed the best performance but in case of cell culture the combination of BAP 2.0 mg/l and 0.2 mg/l NAA showed the best result. Conclusion: The present investigation clearly established and demonstrated the method of obtaining cell suspension culture and important secondary metabolite ricin could be obtained from cell suspension culture of R. communis L. holding promises to explore cell culture industry for ricin production. DOI: http://dx.doi.org/10.3329/jbs.v20i0.17738 J. bio-sci. 20: 161-169, 2012

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Möller, Johannes, and Ralf Pörtner. "Digital Twins for Tissue Culture Techniques—Concepts, Expectations, and State of the Art." Processes 9, no. 3 (March 2, 2021): 447. http://dx.doi.org/10.3390/pr9030447.

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Techniques to provide in vitro tissue culture have undergone significant changes during the last decades, and current applications involve interactions of cells and organoids, three-dimensional cell co-cultures, and organ/body-on-chip tools. Efficient computer-aided and mathematical model-based methods are required for efficient and knowledge-driven characterization, optimization, and routine manufacturing of tissue culture systems. As an alternative to purely experimental-driven research, the usage of comprehensive mathematical models as a virtual in silico representation of the tissue culture, namely a digital twin, can be advantageous. Digital twins include the mechanistic of the biological system in the form of diverse mathematical models, which describe the interaction between tissue culture techniques and cell growth, metabolism, and the quality of the tissue. In this review, current concepts, expectations, and the state of the art of digital twins for tissue culture concepts will be highlighted. In general, DT’s can be applied along the full process chain and along the product life cycle. Due to the complexity, the focus of this review will be especially on the design, characterization, and operation of the tissue culture techniques.

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Goers, Lisa, Paul Freemont, and Karen M. Polizzi. "Co-culture systems and technologies: taking synthetic biology to the next level." Journal of The Royal Society Interface 11, no. 96 (July 6, 2014): 20140065. http://dx.doi.org/10.1098/rsif.2014.0065.

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Co-culture techniques find myriad applications in biology for studying natural or synthetic interactions between cell populations. Such techniques are of great importance in synthetic biology, as multi-species cell consortia and other natural or synthetic ecology systems are widely seen to hold enormous potential for foundational research as well as novel industrial, medical and environmental applications with many proof-of-principle studies in recent years. What is needed for co-cultures to fulfil their potential? Cell–cell interactions in co-cultures are strongly influenced by the extracellular environment, which is determined by the experimental set-up, which therefore needs to be given careful consideration. An overview of existing experimental and theoretical co-culture set-ups in synthetic biology and adjacent fields is given here, and challenges and opportunities involved in such experiments are discussed. Greater focus on foundational technology developments for co-cultures is needed for many synthetic biology systems to realize their potential in both applications and answering biological questions.

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Mišković Špoljarić, Katarina, Marijana Jukić, Teuta Opačak-Bernardi, and Ljubica Glavaš-Obrovac. "3D Cell Technology in Biomedical Research." Collegium antropologicum 44, no. 3 (2020): 171–74. http://dx.doi.org/10.5671/ca.44.3.10.

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Traditional two dimensional cell culture has enabled great strides in biomedicine but needs to be improved to be able to keep up with the demands of modern biomedical research. 2D monolayer culture cannot replicate tissue responses and needs to be supplemented with extensive animal research. Growing cells in three dimensional scaffolds provides a more functional model for biomedical research than traditional monolayer culture. Depending on the needs and the complexity of the model there are several ways that 3D models can be initiated. Simple spheroids can be grown in low adherence plates and in hanging drops while larger spheroids and co-cultured ones need to be grown in systems with greater support such as hydro gels. The system that offers the greatest flexibility is the magnetic levitation approach. In the paper we offer a brief resume to various 3D methods and their characteristics to ease the choice of methods for implementing 3D cell culture techniques.

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Journal articles on the topic 'Cell culture techniques'

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