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Oh, Ji Won, Tsai-Ching Hsi, Christian Fernando Guerrero-Juarez, Raul Ramos, and Maksim V. Plikus. "Organotypic Skin Culture." Journal of Investigative Dermatology 133, no. 11 (November 2013): 1–4. http://dx.doi.org/10.1038/jid.2013.387.
Froeling, Fieke E. M., John F. Marshall, and Hemant M. Kocher. "Pancreatic cancer organotypic cultures." Journal of Biotechnology 148, no. 1 (July 1, 2010): 16–23. http://dx.doi.org/10.1016/j.jbiotec.2010.01.008.
Stahl, Katja, øivind Skare, and Reidun Torp. "Organotypic Cultures as a Model of Parkinson´s Disease. A Twist to an Old Model." Scientific World JOURNAL 9 (2009): 811–21. http://dx.doi.org/10.1100/tsw.2009.68.
Organotypic cultures from the ventral mesencephalon (VM) are widely used to model Parkinson's disease (PD). In this method, neurotoxic compounds have traditionally been applied to the media to induce a uniform dopaminergic (DAergic) cell death in the tissue slices, regardless of the variation existing among slices. This study demonstrates a refinement of the toxic induction technique. We show that unilateral application of 6-hydroxydopamine (6-OHDA) at the tissue surface by means of a microelectrode causes a precisely localized cell death that closely resembles anin vivostereotactic model. This technique introduces an internal control that accounts for variation between slices and enables a precise quantification of the cell loss due to the toxin in use. We characterized organotypic VM cultures in terms of effects of 6-OHDA toxicity and number of DAergic neurons as judged by immunofluorescence and Western blots. Our findings illustrate that this new application technique greatly improves the representativeness of organotypic cultures as a model for PD.We characterized organotypic VM cultures in terms of effects of 6-OHDA toxicity and number of DAergic neurons as judged by immunofluorescence and Western blots. Our findings illustrate that this new application technique greatly improves the representativeness of organotypic cultures as a model for PD.
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Humpel, Christian. "Organotypic Brain Slices of ADULT Transgenic Mice: A Tool to Study Alzheimer’s Disease." Current Alzheimer Research 16, no. 2 (February 4, 2019): 172–81. http://dx.doi.org/10.2174/1567205016666181212153138.
Transgenic mice have been extensively used to study the Alzheimer pathology. In order to reduce, refine and replace (3Rs) the number of animals, ex vivo cultures are used and optimized. Organotypic brain slices are the most potent ex vivo slice culture models, keeping the 3-dimensional structure of the brain and being closest to the in vivo situation. Organotypic brain slice cultures have been used for many decades but were mainly prepared from postnatal (day 8-10) old rats or mice. More recent work (including our lab) now aims to culture organotypic brain slices from adult mice including transgenic mice. Especially in Alzheimer´s disease research, brain slices from adult transgenic mice will be useful to study beta-amyloid plaques, tau pathology and glial activation. This review will summarize the studies using organotypic brain slice cultures from adult mice to mimic Alzheimer's disease and will highlight advantages and also pitfalls using this technique.
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Schuger, L., K. S. O'Shea, B. B. Nelson, and J. Varani. "Organotypic arrangement of mouse embryonic lung cells on a basement membrane extract: involvement of laminin." Development 110, no. 4 (December 1, 1990): 1091–99. http://dx.doi.org/10.1242/dev.110.4.1091.
The behavior of embryonic murine lung cells on a basement membrane extract (Matrigel) was investigated. Single cell suspensions generated by trypsinization of lungs removed from day 12 embryos were plated on Matrigel and cultured for up to one week. The basement membrane extract was used as a gel, and as a wet or dried film. In all of these instances, organotypic arrangement of the embryonic lung cells was observed. This process consisted of cell aggregation, sorting, polarization and formation of a tridimensional organization resembling embryonic lung. The maximal degree of organotypic development was obtained by using a thick gel; minimal reorganization was observed using a dried film. A rabbit polyclonal serum to laminin inhibited organotypic pattern formation while normal rabbit serum did not. Culture of lung cells on laminin gels promoted epithelial cyst formation but poor mesenchymal organization. By studying the behavior of epithelial and/or mesenchymal enriched cell populations on Matrigel, it was concluded that organotypic pattern formation on Matrigel required the presence of both cell populations. Cultivation of dissociated lung cells on a gel consisting of a mixture of collagens type I and III (Vitrogen-100) produced only cell aggregation. Cultivation of lung cells on a thin film of Vitrogen-100 or on uncoated tissue culture plastic produced monolayers of mesenchymal cells alone. Cultivation of lung cells in suspension also failed to induce organotypic arrangement even at maximal cell densities. The present study strongly supports a role for the basement membrane in the organotypic rearrangement of embryonic lung cells and subsequent in vitro cyst formation and budding of the reestablished epithelium. This, in turn, reinforces the concept of the basement membrane as a major regulator of organogenesis.
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Rappoldt, Liam, Adrienne Weeks, Rodney Ouellete, Jeremy Roy, Catherine Taylor, Craig McCormick, Kathleen Attwood, and Inhwa Kim. "TMOD-26. ESTABLISHING A PATIENT-DERIVED, IN-VITRO ORGANOTYPIC SLICE CULTURE MODEL OF GBM." Neuro-Oncology 22, Supplement_2 (November 2020): ii233. http://dx.doi.org/10.1093/neuonc/noaa215.976.
Abstract Glioblastoma Multiforme (GBM) is the most common primary malignant brain tumour. This tumour is universally fatal with a median survival of 15 months. Driving this pathology is an extremely heterogeneic tumour and complex tumour microenvironment. GBM research is primarily conducted using immortalized or primary cell lines due to their practicality and reproducibility. However, these cell lines do not effectively recapitulate the tumour microenvironment. Mouse models address these shortcomings but are laborious and expensive. We propose to utilize a patient derived organotypic culture model of GBM as an intermediary. We have utilized this model to test genetic manipulation via lentiviral transduction and the feasibility of utilizing this model to understand patient derived extracellular vesicles (EVs). We have sectioned and cultured patient derived organotypic models for 14 days without loss of viability. To determine if these organotypic cultures are amenable to lentiviral manipulation, tissue sections were transduced with far-red fluorescent lentivirus and efficiency determined by confocal laser scanning microscopy (CLSM) and flow cytometry (FC). To determine feasibility as a model for EVs, media obtained from patient-derived organotypic cultures was analyzed by western blot, nanoparticle tracking analysis (NTA), and nanoFlow Cytometry (nFC). In the future these EVs will be compared to those found in patient serum. The model of GBM has been lentivirally transduced to express a far-red fluorescent vector in approximately 15% of the slice, quantified by CLSM and FC. EV-sized particles positive for canonical EV markers have been identified in the media by NTA, nFC and western blot. Using lentiviral-mediated genetic engineering and emerging EV science, this organotypic slice culture models yields exciting utility in GBM research. The established organotypic slice culture model will likely be a valuable tool in the study of GBM biology and EV dynamics.
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Ozbun, Michelle A., and Craig Meyers. "Two Novel Promoters in the Upstream Regulatory Region of Human Papillomavirus Type 31b Are Negatively Regulated by Epithelial Differentiation." Journal of Virology 73, no. 4 (April 1, 1999): 3505–10. http://dx.doi.org/10.1128/jvi.73.4.3505-3510.1999.
ABSTRACT Organotypic cultures support the stratification and differentiation of keratinocytes and the human papillomavirus (HPV) life cycle. We report transcription from four novel promoters in the HPV31b upstream regulatory region during the viral life cycle in organotypic cultures. Promoter initiation was not differentiation dependent; two promoters were down-regulated upon epithelial differentiation.
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Brulin, Bénédicte, John C. Nolan, Tecla Marangon, Milan Kovacevic, Mathias Chatelais, Pierre Meheust, Jérome Abadie, et al. "Evaluation of the Chemotherapy Drug Response Using Organotypic Cultures of Osteosarcoma Tumours from Mice Models and Canine Patients." Cancers 13, no. 19 (September 29, 2021): 4890. http://dx.doi.org/10.3390/cancers13194890.
Improvements in the clinical outcome of osteosarcoma have plateaued in recent decades with poor translation between preclinical testing and clinical efficacy. Organotypic cultures retain key features of patient tumours, such as a myriad of cell types organized within an extracellular matrix, thereby presenting a more realistic and personalised screening of chemotherapeutic agents ex vivo. To test this concept for the first time in osteosarcoma, murine and canine osteosarcoma organotypic models were maintained for up to 21 days and in-depth analysis identified proportions of immune and stromal cells present at levels comparable to that reported in vivo in the literature. Cytotoxicity testing of a range of chemotherapeutic drugs (mafosfamide, cisplatin, methotrexate, etoposide, and doxorubicin) on murine organotypic culture ex vivo found limited response to treatment, with immune and stromal cells demonstrating enhanced survival over the global tumour cell population. Furthermore, significantly decreased sensitivity to a range of chemotherapeutics in 3D organotypic culture relative to 2D monolayer was observed, with subsequent investigation confirming reduced sensitivity in 3D than in 2D, even at equivalent levels of drug uptake. Finally, as proof of concept for the application of this model to personalised drug screening, chemotherapy testing with doxorubicin was performed on biopsies obtained from canine osteosarcoma patients. Together, this study highlights the importance of recapitulating the 3D tumour multicellular microenvironment to better predict drug response and provides evidence for the utility and possibilities of organotypic culture for enhanced preclinical selection and evaluation of chemotherapeutics targeting osteosarcoma.
Dissertations / Theses on the topic "Organotypic":
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Li, Daqing. "Entorhino-hippocampal projections in organotypic cultures." Thesis, University College London (University of London), 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.315340.
Tuberculosis predominantly manifests in the form of a pulmonary infection, but may spread out into other parts of the body and is then referred to as extrapulmonary tuberculosis (EPTB). One form of EPTB is an infection of the central nervous system (brain & spinal cord), CNS-TB. Although CNS-TB is relatively rare, accounting for about 5% of EPTB, it is characterised by high morbidity and mortality, particularly for children and immunosuppressed individuals. To examine the effects of a Mycobacterium tuberculosis infection of neural tissue, researchers have hitherto relied on two animal models namely, in vivo intracranial infections or in vitro culturing with dissociated neural cells. Both models have yielded crucial insights concerning CNS-TB but each have limitations e.g. lack of access to the brain during infection in vivo and absence of the 3D organizational tissue structure in vitro. This study investigated the effect of the vaccine strain for tuberculosis, Bacille Calmette-Guerin (BCG) on neural tissue using the model of organotypic hippocampal slice cultures; an in vitro model that overcomes the previously mentioned obstacles. The study sought to expound on immunological and electrophysiological responses to the infection. A viable and moderate BCG infection was established in the hippocampal slice cultures, confirmed by colony forming units enumeration and immunohistochemistry. However, immunological analysis using ELISA found that BCG infection did not change the production levels of cytokines and elicit a distinguishable immune response. To examine the neuronal function during BCG infection, whole-cell patch clamp technique was applied to the hippocampal slice cultures. The neuronal intrinsic properties were not significantly different between infected and non-infected slices. However, tuberculin PPD (M. tuberculosis extract) moderately and transiently had a depolarizing effect when 'puffed' directly onto neurons. In conclusion, organotypic slice cultures are suitable for the investigation of cellular interactions and neural functions in CNS-TB, and the neuronal impact of PPD warrants further investigation.
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Steinmeyer, Joseph D. (Joseph Daly). "Rapid single-cell electroporation for labeling organotypic cultures." Thesis, Massachusetts Institute of Technology, 2010. http://hdl.handle.net/1721.1/60188.
Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2010. Vita. Includes bibliographical references (p. 37-39). Single-cell electroporation is a technique for transfecting individual cells in tissue culture at relatively high efficiencies, however it is both time-consuming and low-throughput and this limits the number of different labeling agents that can be effectively introduced into a region of tissue in reasonable periods of time. A novel system that will rapidly load, clean, and accurately position a glass micropipette electrode into tissue culture for single-cell electroporation is proposed. The system will significantly increase the number of different labeling agents that can be introduced into a single tissue culture per unit time. This in turn, will provide a means for improving the study of neural anatomy at cellular resolutions in both tissue culture and in vivo environments. Supported by grants from the National Institutes of Health and by the MIT Dept. of Electrical Engineering and Computer Science by Joseph D. Steinmeyer. S.M.
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Vu, Lucas Trung. "Proteomic Analysis of Three Dimensional Organotypic Liver Models." Diss., Virginia Tech, 2015. http://hdl.handle.net/10919/77033.
In vitro liver models that closely mimic the in vivo microenvironment are central for understanding hepatic functions and intercellular communication processes. Bottom-up shotgun proteomic analysis of the hepatic cells can lend insight into such processes. This technique employs liquid chromatography-tandem mass spectrometry (LC-MS/MS) for relative quantification of protein abundances by measuring intensities of their corresponding peptides. Organotypic 3D liver models have been developed in our laboratory that consist of hepatocytes and liver sinusoidal endothelial cells (LSECs) separated by a polyelectrolyte multilayer (PEM), which serves as a mimic for the Space of Disse. Each component within these models is easily separable allowing for systematic evaluation of the cells and PEMs. In this study, proteomes of hepatocytes from PEM containing models, cultured with and without LSECs, were compared to those from monolayers. Changes in core metabolism were evaluated among all culture conditions. Overall, all cultures were ketogenic and performed gluconeogenesis. The presence of the PEM led to increases in proteins associated with mitochondrial-based β-oxidation and peroxisomal proteins. The PEMs also limited production of structural proteins, which are linked to dedifferentiation of hepatocytes, suggesting that cell-ECM interactions are essential for maintenance of their liver-like state. The presence of LSECs increased levels of carboxylesterases and other phase I and phase II detoxification enzymes suggesting that intercellular signaling mediates enzyme abundance. Taken together, these results suggest that the cell-cell (from the LSECs) and cell-ECM (from the PEMs) interactions exert different, yet crucial effects, and both are required for the preservation of metabolic liver functions and differentiated phenotypes. Changes in the PEMs as a result of cell culture were also evaluated but exhibited minimal differences at this time point. Proteomes of LSECs monolayers were also characterized. Enzymes related to the metabolism of amino acids, lipids, oxidative phosphorylation and phase I and phase II detoxification processes were all identified in LSECs monolayers highlighting their role in these processes. Characterization of 3DHL LSECs was not possible due to ion suppression resulting from the presence of excess contaminant proteins. Nonetheless, this study provides a foundation in which LSECs from 3D liver models can be compared against in future studies. Ph. D.
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Kazmi, B. "Oral and dermal fibroblasts in 3D organotypic co-culture." Thesis, University College London (University of London), 2009. http://discovery.ucl.ac.uk/17485/.
It is widely observed that dermal wound healing results in the re-establishment of skin continuity through the formation of a scar. In comparison oral wounds heal more rapidly, with less visible scarring. The diverse nature of sub-epithelial fibroblasts has targeted them as mediators for such a distinction in phenotype. Fibroblasts and their differentiated form, the myofibroblast (characterised by the expression of α-SMA), are chief synthesisers of the ECM during wound healing. Myofibroblasts in particular play an important role in wound contracture, ECM synthesis and remodelling during normal wound healing and scar production; but are also observed in fibrosis and pathological scarring. The most potent mediator of this phenotype in the cytokine TGFβ1, which is abundantly present throughout wound healing. Here we used organotypic co-cultures and mono-cultures to assess the differences in phenotype and the relative contribution of topographically distinct fibroblasts within them. There is currently little data on the responsiveness of oral buccal mucosal and skin fibroblasts to TGFβ1, particularly when keratinocytes are present in a 3D environment. Here, for the first time, we used heterotypic organotypic co-cultures containing skin and oral buccal mucosal fibroblasts and added TGFβ1 under both resting and tethered conditions. We monitored the changes in response of these fibroblasts with and without the presence of keratinocytes, and subsequently assessed changes in phenotype upon substitution of the dermal fibroblasts with those from the reduced scarring oral buccal mucosa. Under resting conditions oral fibroblast seeded organotypics showed a lower constitutive myofibroblast expression, and were unresponsive to TGFβ1 mediated α- SMA expression, regardless of substrate compliance. These fibroblasts were found to significantly promote epithelial maturation under resting conditions, and express higher levels of active MMP-9 (and enzyme involved with keratinocyte migration) upon treatment with TGFβ1, when compared to dermal fibroblast seeded counterparts. When oral fibroblasts were introduced into a dermal organotypic co-culture environment, we observed a preferential change α-SMA phenotype, offering the potential for use of these cells in an area distinct from there origin to a favourable effect.
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Frantseva, Marina. "H¦2O¦2 induced toxicity in rat organotypic hippocampal cultures." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape11/PQDD_0028/MQ51596.pdf.
Breathing is an important rhythmic motor behavior whose underlying neural mechanisms can be studied in vitro. The study of breathing rhythms in vitro has depended upon reduced preparations of the brainstem that both retain respiratory-active neuronal populations and spontaneously generate respiratory-related motor output from cranial and spinal motor nerves. Brainstem-spinal cord en bloc preparations and transverse medullary slices of the brainstem have greatly improved the ability of researchers to experimentally access and thus characterize interneurons important in respiratory rhythmogenesis. These existing in vitro preparations are, however, not without their limitations. For example, the window of time within which experiments may be conducted is limited to several hours. Moreover, these preparations are poorly suited for studying subcellular ion channel distributions and synaptic integration in dendrites of rhythmically active respiratory interneurons because of tortuous tissue properties in slices and en bloc, which limits imaging approaches. Therefore, there is a need for an alternative experimental approach. Acute transverse slices of the medulla containing the preBötzinger complex (preBötC) have been exploited for the last 25 years as a model to study the neural basis of inspiratory rhythm generation. Here we transduce such preparations into a novel organotypic slice culture that retains bilaterally synchronized rhythmic activity for up to four weeks in vitro. Properties of this culture model of inspiratory rhythm are compared to analogous acute slice preparations and the rhythm is confirmed to be generated by neurons with similar electrophysiological and pharmacological properties. The improved optical environment of the cultured brain tissue permits detailed quantitative calcium imaging experiments, which are subsequently used to examine the subcellular distribution of a transient potassium current, IA, in rhythmically active preBötC interneurons. IA is found on the dendrites of these rhythmically active neurons, where it influences the electrotonic properties of dendrites and has the ability to counteract depolarizing inputs, such as post-synaptic excitatory potentials, that are temporally sparse in their occurrence (i.e., do not summate). These results suggest that excitatory input can be transiently inhibited by IA prior to its steady-state inactivation, which would occur as temporally and spatially summating synaptic inputs cause persistent depolarization. Thus, rhythmically active interneurons are equipped to appropriately integrate the activity state of the inspiratory network, inhibiting spurious inputs and yet yielding to synaptic inputs that summate, which thus coordinates the orderly recruitment of network constituents for rhythmic inspiratory bursts. In sum, the work presented here demonstrates the viability and potential usefulness of a new experimental model of respiratory rhythm generation, and further leverages its advantages to answer questions about dendritic synaptic integration that could not previously be addressed in the acute slice models of respiration. We argue that this new organotypic slice culture will have widespread applicability in studies of respiratory rhythm generation.
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Jussila, T. (Tommi). "Modelling cancer: recapitulation of tumor growth in experimental systems in vivo and in vitro." Doctoral thesis, University of Oulu, 2000. http://urn.fi/urn:isbn:9514256433.
Abstract
The purpose of the study was to evaluate model systems of cancer development and
compare some of their critical features with cancer development in
vivo. Ovarian and endometrial cancers in man were used as correlates.
Tumor development in experimental animals, exposed to carcinogens and UV
irradiation, showed the entire spectrum of tumor development as compared to
spontaneous carcinomas: hyperplasia, dysplasia, benign papillomas and malignant
squamous cell carcinomas. For short-term analysis of differentiated homogenous
cell populations, the transplant model proved most useful. For long term
analysis of effects of extraneous agents, the skin carcinogenesis model is
probably the most rewarding.
Analysis of proliferation markers in human tumor samples as studied by
immunohistochemistry, showed that an increased expression of PCNA and Ki-67 was
associated with poor prognosis in ovarian neoplasms. Analysis of cell
proliferation in model tumors showed that the transplant model has a better
sensitivity when compared to the animal skin model and the subcutaneous
injection model, in that effect of changes in cell-host interaction on the
location and extent of the proliferating cell population can be studied therein.
The expression of some growth factors, their receptors, oncogenes and suppressor
genes were studied in ovarian and endometrial carcinomas and in skin cancer
model system in mouse exposed to carcinogens and UV irradiation. Variability in
expression and methodological problems precluded detailed analysis of these
markers in different models.
The expression of TGFβ1, TGFβ2 and TGFβ3 was determined in normal
human keratinocytes, and in 7 immortalized and
ras-transfected benign and malignant keratinocyte cell
lines, maintained as transplants and as subcutaneous tumors in nude mice. By
differential immunohistochemical localization of TGFβ isoforms, we
demonstrated that each isoform may serve specific roles in tumor development and
progression. The complex nature of TGFβ expression prevented detailed
analysis of isozymes in different models, the results in this study, however,
indicated a similar pattern in the models analyzed.
Morphological methods were used to determine relationship between epithelial
growth and formation and deposition of collagens in the extracellular matrix in
experimental models and human tumors. The composition of the mesenchyme differed
in tumors originating from different cell lines reflecting functional
interaction between epithelial cells and the mesenchyme in neoplastic
development. Tumor-stroma interaction was distinct in human, comparable
alterations were observed in experimental models, more so in transplants, less
in subcutaneous tumors, affecting tumor growth and differentiation in the
different models.
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Acheva, Anna Rumenova. "Mechanisms of response to targeted irradiation in organotypic 3D skin cultures." Thesis, Queen's University Belfast, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.579565.
Low linear energy transfer (LET) ionizing radiation as gamma and X-rays is widely used in the modem medicine for diagnostic and treatment purposes. Despite all the advantages that it gives for the early diagnosis and in the radiotherapy treatment of cancer, there are still unclear aspects about the mechanisms of the radiation effects in human tissue. Of particular interest is the detailed pathway which the directly irradiated cells use to communicate to their neighbours and its possible implications for radiotherapy applications. The aim of this project is to study the spatio-temporal signalling from irradiated to non- irradiated cells using in vitro 3D tissue models. For evaluation of radiation induced effects we used conventional 225 kVp X-ray and lead shielding to observe the effects in the non-targeted cells. Additionally we applied the novel 30 kVp micro collimator that could irradiate samples in wide 1-10 urn lines. The application of this targeted radiation source together with the shielding irradiation set up will cast light on the effects induced by localized radiation exposures and the signalling from the irradiated to non-irradiated cells using 3D organotypic skin as a model. Due to its fast cell turnover, the human epidermis is extremely sensitive to IR. This has a limiting effect on the radiotherapy as severe normal skin responses can delay treatment and even decline patients from radiotherapy. The current work is aiming to reveal the mechanisms of DNA damage, repair and their later consequences for the differentiation and development of inflammatory-like response in the irradiated and surrounding areas within the 3D organotypic skin model. We investigated use of inhibitors of pro-inflammatory pathways such as the transcription factor NF-KB and its downstream target COX-2, in order to reduce the signal transduction from the irradiated to non irradiared cells and to reduce the inflammatory responses in the surrounding normal tissue. Similar methods of control of the signal diffusion could be applied in the radiotherapy to reduce the inflammatory skin responses and decrease the range of the late side effects developing from the chronic inflammation that could further lead to ulceration and skin necrosis.
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Froeling, Fieke E. M. "Tumour–stroma interactions in an organotypic culture model of pancreatic cancer." Thesis, Queen Mary, University of London, 2011. http://qmro.qmul.ac.uk/xmlui/handle/123456789/705.
Pancreatic cancer is characterised by an intense fibrotic, or desmoplastic, stroma, which contributes to tumour progression. Three-dimensional in vitro culture models incorporating this non-tumour component may more closely recapitulate the complex in vivo situation. The aim of my project was to develop a physiologically relevant, three-dimensional organotypic culture model of pancreatic cancer to study the tumour-stroma interactions and its modulation by novel therapeutic agents. Cancer cells cultured on top of collagen/Matrigel gels, embedded with or without stromal cells (hTERT immortalised PS1 stellate cells or MRC5 fibroblasts), differentiated into luminal structures, exhibiting a central apoptotic core with a proliferating peripheral rim and apicobasal polarity. Stromal cells induced a reduction in total tumour cell number, which was associated with a decrease in E-cadherin expression, upregulated β-catenin expression and translocation of ezrin from the apical to the basal aspect of cancer cells, where it was associated with invasive activity. Subsequently, this organotypic model was raised to an air-liquid interface to study the direct and indirect effects of all-trans retinoic acid (ATRA), which rendered stellate cells back to their quiescent phenotype. Indirect effects of quiescent stellate cells on pancreatic cancer cells included changes in proliferation (decrease), apoptosis (increase), invasion (decrease), Wnt/β-catenin signalling (decrease) and an altered morphology. The Wnt/β-catenin signalling 6 perturbations were mediated by restoration of sFRP4 (secreted frizzled-related protein 4) secretion by quiescent stellate cells, resulting in reduced cancer cell invasion (reporter and invasion assays). All such observations could be validated in human pancreatic cancer tissue samples. Taken together, pancreatic organotypic culture offers a reproducible, in vitro three-dimensional culture model, which allows the study of tumour-stroma interactions in a physiologically relevant system. For treatment of pancreatic cancer, a tumour characterised by a poor response to conventional chemotherapeutic drugs, targeting the tumour-stroma cross-talk with agents such as ATRA offers an exciting novel therapeutic strategy.
Schäfer-Korting, Monika, Silvya Stuchi Maria-Engler, and Robert Landsiedel, eds. Organotypic Models in Drug Development. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-70063-8.
Bagnoli, Fabio, and Rino Rappuoli, eds. Three Dimensional Human Organotypic Models for Biomedical Research. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-62452-1.
Schäfer-Korting, Monika, Robert Landsiedel, and Silvya Stuchi Maria-Engler. Organotypic Models in Drug Development. Springer International Publishing AG, 2021.
Schäfer-Korting, Monika, Robert Landsiedel, and Silvya Stuchi Maria-Engler. Organotypic Models in Drug Development. Springer International Publishing AG, 2022.
Pasonen-Seppanen, Sanna. REGULATION OF KERATINOCYTE DIFFERENTIATION AND HYALURONAN METABOLISM IN AN ORGANOTYPIC KERATINOCYTE CULTURE [KUOPIO UNIVERSITY PUBLIATIONS D. MEDICAL SCIENCES, 370]. Kuopion Yliopisto Kuopio, Finland, 2005.
Li, Zhong, Shiqi Xiang, Eileen N. Li, Madalyn R. Fritch, Peter G. Alexander, Hang Lin, and Rocky S. Tuan. "Tissue Engineering for Musculoskeletal Regeneration and Disease Modeling." In Organotypic Models in Drug Development, 235–68. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/164_2020_377.
Marionnet, Claire, and Françoise Bernerd. "Organotypic Models for Evaluating Sunscreens." In Principles and Practice of Photoprotection, 199–225. Cham: Springer International Publishing, 2016. http://dx.doi.org/10.1007/978-3-319-29382-0_12.
Müller, Brigitte. "Organotypic Culture of Adult Mouse Retina." In Methods in Molecular Biology, 181–91. New York, NY: Springer New York, 2019. http://dx.doi.org/10.1007/978-1-4939-9086-3_13.
Jenei, Veronika, Maria L. Nystrom, and Gareth J. Thomas. "Measuring Invasion in an Organotypic Model." In Methods in Molecular Biology, 223–32. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-207-6_15.
Zhu, Lucía, and Manuel Valiente. "Organotypic Brain Cultures for Metastasis Research." In Brain Tumors, 119–32. New York, NY: Springer US, 2020. http://dx.doi.org/10.1007/978-1-0716-0856-2_6.
Allen, Jennifer L., and Harry Mellor. "The Coculture Organotypic Assay of Angiogenesis." In Methods in Molecular Biology, 265–70. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-1462-3_17.
Hofmann, Hans-Dieter, Steffen Schulz-Key, Daniel Hertle, and Matthias Kirsch. "Organotypic Cultures of the Rat Retina." In New Methods for Culturing Cells from Nervous Tissues, 58–73. Basel: KARGER, 2005. http://dx.doi.org/10.1159/000083442.
Batourina, Ekaterina, Devangini Gandhi, Cathy L. Mendelsohn, and Andrei Molotkov. "Organotypic Culture of the Urogenital Tract." In Kidney Development, 45–53. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-851-1_5.
Dash-Wagh, Suvarna, Ülo Langel, and Mats Ulfendahl. "PepFect6 Mediated SiRNA Delivery into Organotypic Cultures." In Methods in Molecular Biology, 27–35. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-3112-5_3.
Sundstrom, Lars E., Igor Charvet, and Luc Stoppini. "Organotypic microtissues on an air-liquid interface." In Technology Platforms for 3D Cell Culture, 123–42. Chichester, UK: John Wiley & Sons, Ltd, 2017. http://dx.doi.org/10.1002/9781118851647.ch6.
Shull, Gabriella, Christiane Haffner, Wieland Huttner, Elena Taverna, and Suhasa B. Kodandaramaiah. "Robotic Platform for the Delivery of Gene Products Into Single Cells in Organotypic Slices of the Developing Mouse Brain." In 2018 Design of Medical Devices Conference. American Society of Mechanical Engineers, 2018. http://dx.doi.org/10.1115/dmd2018-6899.
Microinjection of genetic components and dye into organotypic slices provides excellent single cell resolution for unraveling biological complexities, but is extremely difficult and time consuming to perform manually resulting in low yield and low use in the developmental biology field. We developed a computer vision guided platform to inject specimen with mRNA, and/or dye and investigated the efficiency of the process using organotypic slices of the mouse developing neocortex. We demonstrate that the system significantly increases yield of injection relative to manual use by an order of magnitude, allows for cell tracking over 0, 24, and 48 hours post injection in culture, and enables mRNA translation of injected product. The autoinjector platform thus can open the door to new types of experiments including investigating effects of mRNA concentration, and composition on cell fate, and tracking these effects on cell reprogramming and lineage.
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Dao, Belinda, Alissa Yamazaki, Chung Ho Sun, Zifu Wang, Nguyen Pham, Michael Oldham, and Brian J. F. Wong. "Multi-photon microscopy of tobacco-exposed organotypic skin models." In Biomedical Optics 2006, edited by Nikiforos Kollias, Haishan Zeng, Bernard Choi, Reza S. Malek, Brian J. Wong, Justus F. R. Ilgner, Eugene A. Trowers, et al. SPIE, 2006. http://dx.doi.org/10.1117/12.674185.
Sprackling, Carley M., Jeremy D. Kratz, Peter F. Favreau, Mohammad R. Karim, Christopher P. Babiarz, Cheri A. Pasch, Amani A. Gillette, et al. "Abstract 3143: Predicting treatment response using patient derived organotypic cancer spheroids." In Proceedings: AACR Annual Meeting 2019; March 29-April 3, 2019; Atlanta, GA. American Association for Cancer Research, 2019. http://dx.doi.org/10.1158/1538-7445.am2019-3143.
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