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Journal articles on the topic 'Hepatocyte isolation'

Author: Grafiati

Published: 4 June 2021

Last updated: 12 February 2022

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1

C. S., Sushrutha, Sandhya K., Savitha Karalwad, and Elango E. M. "Recommended parameters for hepatocyte isolation-a review." International Journal of Scientific Reports 6, no. 12 (2020): 537. http://dx.doi.org/10.18203/issn.2454-2156.intjscirep20205035.

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<p>There is an increasing demand for primary human hepatocytes for toxico-pharmacological studies and for exploring their role in liver cell transplantation. During the whole process of isolation, culture and cryopreservation hepatocytes are subjected to various stress. All these factors will lead to the impairment in the metabolic capability of the hepatocytes post isolation. Hepatocyte isolation is an expensive and resource driven procedure. Thus, it is important to consider all the factors before we subject the specimen for isolation of cells. By considering the factors that influence

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2

Chen, Ya-Hui, Hui-Ling Chen, Cheng-Maw Ho, et al. "Preclinical Application of Reduced Manipulated Processing Strategy to Collect Transplantable Hepatocytes: A Pilot and Feasibility Study." Journal of Personalized Medicine 11, no. 5 (2021): 326. http://dx.doi.org/10.3390/jpm11050326.

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Background: The complex isolation and purification process of hepatocytes for transplantation is labor intensive and with great contamination risk. Here, as a pilot and feasibility study, we examined in vitro and in vivo hepatocyte isolation feasibility and cell function of Cell Saver® Elite®, an intraoperative blood-cell-recovery system. Methods: Rat and pig liver cells were collected using this system and then cultured in vitro, and their hepatocyte-specific enzymes were characterized. We then transplanted the hepatocytes in an established acute liver–injured (retrorsine+D-galactosamine-trea

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3

Nieuwoudt, M. J., E. Kreft, B. Olivier, et al. "A Large-Scale Automated Method for Hepatocyte Isolation: Effects on Proliferation in Culture." Cell Transplantation 14, no. 5 (2005): 291–99. http://dx.doi.org/10.3727/000000005783983007.

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Large-scale sterile methods for isolating hepatocytes are desirable for the development of bioartificial liver support systems. In this study the traditional centrifuge method was compared with the use of a Baylor Rapid Autologous Transfusion (BRAT) machine for isolating large quantities of porcine hepatocytes. After isolating hepatocytes, the methods were evaluated in terms of cell viability and yield per liver, proliferation over 7 days, and the effects on the cell cycle using the trypan blue exclusion test, conventional phase-contrast light microscopy, the lactate to pyruvate ratio, the lea

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4

Lehec, Sharon C., Robin D. Hughes, Ragai R. Mitry, et al. "Experience of Microbiological Screening of Human Hepatocytes for Clinical Transplantation." Cell Transplantation 18, no. 8 (2009): 941–47. http://dx.doi.org/10.3727/096368909x471323.

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Hepatocyte transplantation is being used in patients with liver-based metabolic disorders and acute liver failure. Hepatocytes are isolated from unused donor liver tissue under GMP conditions. Cells must be free of microbiological contamination to be safe for human use. The experience of microbiological screening during 72 hepatocyte isolation procedures at one center is reported. Samples were taken at different stages of the process and tested using a blood culture bottle system and Gram stain. Bacterial contamination was detected in 37.5% of the UW organ preservative solutions used to transp

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5

Kobayashi, Naoya, and Noriaki Tanaka. "Engineering of Human Hepatocyte Lines for Cell Therapies in Humans: Prospects and Remaining Hurdles." Cell Transplantation 11, no. 5 (2002): 417–20. http://dx.doi.org/10.3727/000000002783985693.

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Hepatocyte-based biological therapies are increasingly envisioned for temporary support in acute liver failure and provision of specific-liver functions in liver-based metabolic deficiency. One of the hurdles to develop such therapies is severe shortage of human livers for hepatocyte isolation. To address the issue, we have focused on reversible immortalization of human hepatocytes. Such technology can allow rapid preparation of functional and uniform human hepatocytes. Here we present our strategy to construct transplantable human hepatocyte cell lines.

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6

Tolosa, Laia, Eugenia Pareja-Ibars, M. Teresa Donato, et al. "Neonatal Livers: A Source for the Isolation of Good-Performing Hepatocytes for Cell Transplantation." Cell Transplantation 23, no. 10 (2014): 1229–42. http://dx.doi.org/10.3727/096368913x669743.

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Hepatocyte transplantation is an alternative therapy to orthotopic liver transplantation for the treatment of liver diseases. However, the supply of hepatocytes is limited given the shortage of organs available to isolate good-functioning quality cells. Neonatal livers may be a potential source alternative to adult livers to obtain good-performing hepatic cells for hepatocyte transplantation, which has not yet been explored profoundly. High-yield preparations of viable hepatocytes were isolated from 1- to 23-day-old liver donors, cryopreserved, and banked. Cell integrity and functional quality

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7

Dhanasekaran, Sugapriya, Devilakshmi Sithambaram, Kavitha Govarthanan, Bijesh Kumar Biswal, and Rama S. Verma. "An Efficient Protocol for Deriving Liver Stem Cells from Neonatal Mice: Validating Its Differentiation Potential." Analytical Cellular Pathology 2015 (2015): 1–10. http://dx.doi.org/10.1155/2015/219206.

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The success of liver regeneration depends on the availability of suitable cell types and their potential to differentiate into functional hepatocytes. To identify the stem cells which have the ability to differentiate into hepatocytes, we used neonatal liver as source. However, the current protocol for isolating stem cells from liver involves enzymes like collagenase, hyaluronidase exposed for longer duration which limits the success. This results in the keen interest to develop an easy single step enzyme digestion protocol for isolating stem cells from liver for tissue engineering approaches.

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8

Montanari, Elisa, Joel Pimenta, Luca Szabó, et al. "Beneficial Effects of Human Mesenchymal Stromal Cells on Porcine Hepatocyte Viability and Albumin Secretion." Journal of Immunology Research 2018 (2018): 1–13. http://dx.doi.org/10.1155/2018/1078547.

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Porcine hepatocytes transplanted during acute liver failure might support metabolic functions until the diseased liver recovers its function. Here, we isolated high numbers of viable pig hepatocytes and evaluated hepatocyte functionality after encapsulation. We further investigated whether coculture and coencapsulation of hepatocytes with human multipotent mesenchymal stromal cells (MSC) are beneficial on hepatocyte function. Livers from 10 kg pigs (n=9) were harvested, and hepatocytes were isolated from liver suspensions for microencapsulation using alginate and poly(ethylene-glycol)- (PEG-)

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9

Sato, Masahiro, Issei Saitoh, Emi Inada, Shingo Nakamura, and Satoshi Watanabe. "Potential for Isolation of Immortalized Hepatocyte Cell Lines by Liver-DirectedIn VivoGene Delivery of Transposons in Mice." Stem Cells International 2019 (June 2, 2019): 1–12. http://dx.doi.org/10.1155/2019/5129526.

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Isolation of hepatocytes and their culturein vitrorepresent important avenues to explore the function of such cells. However, these studies are often difficult to perform because of the inability of hepatocytes to proliferatein vitro. Immortalization of isolated hepatocytes is thus an important step toward continuousin vitroculture. For cellular immortalization, integration of relevant genes into the host chromosomes is a prerequisite. Transposons, which are mobile genetic elements, are known to facilitate integration of genes of interest (GOI) into chromosomesin vitroandin vivo. Here, we prop

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10

Yin, Zhaohui, Ewa C. S. Ellis, and Greg Nowak. "Isolation of Mouse Hepatocytes for Transplantation: A Comparison between Antegrade and Retrograde Liver Perfusion." Cell Transplantation 16, no. 8 (2007): 859–65. http://dx.doi.org/10.3727/000000007783465235.

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We compared antegrade with retrograde liver perfusion when isolating mouse hepatocytes for hepatocyte transplantation. Male mouse hepatocytes were isolated by different perfusion methods and transplanted into the spleen of congeneic female mice. Retrograde perfusion yielded a larger number of cells (4.90 × 107) than antegrade (4.09 × 107, p < 0.05), but hepatocytes obtained by antegrade perfusion gave higher engraftment efficiency (p < 0.05). More of the transplanted hepatocytes could be recovered from recipient liver with antegrade perfusion than with retrograde perfusion (p < 0.05).

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11

Hendrawan, Siufui, Jennifer Lheman, Nuraeni, Ursula Weber, and Hans Ulrich Baer. "Hepatocyte and Islet Cell Cotransplantation on Poly-L-Lactide Matrix for the Treatment of Liver Cirrhosis." International Journal of Hepatology 2020 (October 13, 2020): 1–6. http://dx.doi.org/10.1155/2020/5410359.

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The human autologous hepatocyte matrix implant is a promising alternative procedure to counter liver damage. We assessed the outcome of human hepatocytes isolation from cirrhotic liver compared to the clinical and histological scores of disease severity. A total of 11 patients with various clinical scores (CTP and MELD) and histological score (Metavir, fibrosis) of liver cirrhosis were included in the hepatocyte matrix implant clinical phase I study. The liver segment and pancreatic tissue were harvested from each patient, and hepatocytes and cells of islets of Langerhans were isolated. The fr

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12

Donato, María Teresa, Agustín Lahoz, Sandra Montero, et al. "Functional Assessment of the Quality of Human Hepatocyte Preparations for Cell Transplantation." Cell Transplantation 17, no. 10-11 (2008): 1211–19. http://dx.doi.org/10.3727/096368908787236620.

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Hepatocyte transplantation is an alternative therapy to orthotopic liver transplantation for the treatment of liver diseases. Good quality freshly isolated or cryopreserved human hepatocytes are needed for clinical transplantation. However, isolation, cryopreservation, and thawing processes can seriously impair hepatocyte viability and functionality. The aim of the present study was to develop a fast and sensitive procedure to estimate the quality of hepatocyte preparations prior to clinical cell infusion. To this end, cell viability, attachment efficiency, and metabolic competence (urea synth

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13

Alexandrova, Krassimira, Carsten Griesel, Marc Barthold, et al. "Large-Scale Isolation of Human Hepatocytes for Therapeutic Application." Cell Transplantation 14, no. 10 (2005): 845–53. http://dx.doi.org/10.3727/000000005783982530.

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During the last decade, hepatocyte transplantation has been suggested as a safe and potentially effective clinical option for the treatment of acute or decompensating chronic liver failure as well as for hereditary liver disease. Currently, one of the major limiting factors for clinical application is the insufficient access to suitable liver cell preparations. In cooperation with the German and Catalane organ procurement organizations, a routine procedure for the isolation of hepatocytes from donor organs rejected for transplantation (n = 117) has been established. The process is performed ac

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14

Horner, Rosa, Martin Kluge, Joseph Gassner, et al. "Hepatocyte Isolation After Laparoscopic Liver Resection." Tissue Engineering Part C: Methods 22, no. 9 (2016): 839–46. http://dx.doi.org/10.1089/ten.tec.2016.0187.

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15

Charni-Natan, Meital, and Ido Goldstein. "Protocol for Primary Mouse Hepatocyte Isolation." STAR Protocols 1, no. 2 (2020): 100086. http://dx.doi.org/10.1016/j.xpro.2020.100086.

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16

Ambrosino, Giovanni, Stefano M. M. Basso, Sergio Varotto, Enrico Zardi, Antonio Picardi, and Davide F. D'amico. "Isolated Hepatocytes versus Hepatocyte Spheroids: In Vitro Culture of Rat Hepatocytes." Cell Transplantation 14, no. 6 (2005): 397–401. http://dx.doi.org/10.3727/000000005783982954.

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The use of hepatocytes that express liver-specific functions to develop an artificial liver is promising. Unfortunately, the loss of specialized liver functions (dedifferentiation) is still a major problem. Different techniques, such as collagen entrapment, spherical multicellular aggregates (spheroids), and coculture of hepatocytes with extracellular matrix, have been used to improve the performance of hepatocytes in culture. The aim of this study was to compare two different models of hepatocyte isolation in culture: isolated hepatocytes (G1) and hepatocyte spheroids (60% hepatocytes, 40% no

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17

Moon, T. W., P. J. Walsh, and T. P. Mommsen. "Fish Hepatocytes: A Model Metabolic System." Canadian Journal of Fisheries and Aquatic Sciences 42, no. 11 (1985): 1772–82. http://dx.doi.org/10.1139/f85-222.

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The hepatocyte or liver cell preparation is a standard metabolic model in mammalian physiology/biochemistry. This paper presents a basic method for the isolation of viable fish hepatocytes, reviews specifically the literature available on hepatic function and adaptation using the preparation, and examines those areas where this preparation could contribute to our understanding of basic and applied fisheries biology. Viable liver cells are prepared by collagenase perfusion and collected by low-speed centrifugation. Buffered salines employed for cell isolation must be consistent with the normal

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18

Mitry, Ragai R., Robin D. Hughes, Marion M. Aw, et al. "Human Hepatocyte Isolation and Relationship of Cell Viability to Early Graft Function." Cell Transplantation 12, no. 1 (2003): 69–74. http://dx.doi.org/10.3727/000000003783985197.

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Hepatocyte transplantation is emerging as an additional modality of treatment for patients with acute liver failure or liver-based metabolic disorders. The procedure requires isolation of high-quality hepatocytes from unused donor livers. Hepatocytes were isolated from 20 donor livers (11 right lobes, 3 left lateral segments, 6 whole livers) using a collagenase perfusion technique. Cell viability (median 56%, range 13–95%) and yield (median 1.4 × 109 cells, range 2.0 × 106–1.8 × 1010 cells) varied according to the tissue available. Fatty livers rejected for transplantation gave lower cell viab

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19

Lee, Doo-Hoon, and Kwang-Woong Lee. "Hepatocyte Isolation, Culture, and Its Clinical Applications." Hanyang Medical Reviews 34, no. 4 (2014): 165. http://dx.doi.org/10.7599/hmr.2014.34.4.165.

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20

Maruyama, Masanobu, Toshinori Totsugawa, Takemi Kunieda, et al. "Hepatocyte Isolation and Transplantation in the Pig." Cell Transplantation 12, no. 6 (2003): 593–98. http://dx.doi.org/10.3727/000000003108747190.

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21

Klaunig, James E., Randall J. Ruch, and Peter J. Goldblatt. "Trout hepatocyte culture: Isolation and primary culture." In Vitro Cellular & Developmental Biology 21, no. 4 (1985): 221–28. http://dx.doi.org/10.1007/bf02620933.

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22

CORNO. "Human hepatocyte isolation from rejected whole organs." Cell Transplantation 5, no. 5 (1996): 8. http://dx.doi.org/10.1016/0963-6897(96)82091-4.

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23

Terry, Claire, Ragai R. Mitry, Sharon C. Lehec, et al. "The Effects of Cryopreservation on Human Hepatocytes Obtained from Different Sources of Liver Tissue." Cell Transplantation 14, no. 8 (2005): 585–94. http://dx.doi.org/10.3727/000000005783982765.

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Successful cryopreservation of human hepatocytes is important to establish hepatocyte banks for clinical use or in vitro research. The availability of donor tissue from unused liver segments/lobes and non-heart-beating donors (NHBD) has provided newer sources of hepatocytes. The quality of hepatocytes at the time of cryopreservation is important as cells isolated from liver tissue of borderline quality may not withstand the stresses associated with cryopreservation and subsequent thawing. Human hepatocytes were cryopreserved after isolation from mainly donor tissues (n = 40). In vitro assessme

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24

Kobayashi, Naoya, Masahiro Miyazaki, Kenichi Fukaya, et al. "Treatment of Surgically Induced Acute Liver Failure with Transplantation of Highly Differentiated Immortalized Human Hepatocytes." Cell Transplantation 9, no. 5 (2000): 733–35. http://dx.doi.org/10.1177/096368970000900524.

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Primary human hepatocytes are an ideal source of hepatic function in bioartficial liver (BAL), but the shortage of human livers available for hepatocyte isolation limits this modality. To resolve this issue, primary human fetal hepatocytes were immortalized using simian virus 40 large T antigen. One of the immortal cell lines, OUMS-29, showed highly differentiated liver functions. Intrasplenic transplantation of OUMS-29 cells protected 90% hepatectomized rats from hyperammonemia and significantly prolonged their survival. Essentially unlimited availability of OUMS-29 cells supports their clini

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25

Al Battah, Feras, Joery De Kock, Tamara Vanhaecke, and Vera Rogiers. "Current Status of Human Adipose–Derived Stem Cells: Differentiation into Hepatocyte-Like Cells." Scientific World JOURNAL 11 (2011): 1568–81. http://dx.doi.org/10.1100/tsw.2011.146.

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The shortage of human organ donors and the low cell quality of available liver tissues represent major obstacles for the clinical application of orthotropic liver transplantation and hepatocyte transplantation, respectively. Therefore, worldwide research groups are investigating alternative extrahepatic cell sources. Recentin vitrostudies have demonstrated that mesenchymal stem cells (MSCs) from various sources, including human bone marrow, adipose tissue, and umbilical cord, can be differentiated into hepatocyte-like cells when appropriate conditions are used. In particular, interest exists f

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26

Nguyen, Ngoc Hong, Giau Van Vo, Thuy Ngoc Huynh, Hung Tran, and Duong Huynh Thuy Ho. "HEPATOCYTE ISOLATION AND EX VIVO HEPATOPROTECTION OF ORTHOSIPHON ARISTATUS EXTRACT ON CARBON TETRACHLORIDE INDUCED HEPATOTOXICITY." Science and Technology Development Journal 13, no. 3 (2010): 58–66. http://dx.doi.org/10.32508/stdj.v13i3.2154.

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Orthosiphon aristatus Blume, a member of the Lamiaceac family, is a medicinal herb known useful as a diuretic agent, used popularly in Vietnam and the nations in the world. In our previous research, it is found that O. aristatus had strong antioxidant in both methods of the ferric reducing/antioxidant power assay and the 1,1-diphenyl-2-picryl-hydrazyl free radical scavenging assay. The purpose of the present study was to examine the methanolic extract of O. aristatus with strong antioxidative activity against carbon tetrachloride (CCl4) induced cytotoxicity in isolated mouse hepatocytes using

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27

Levitsky, L. L., Q. Zheng, K. Mink, and D. B. Rhoads. "GLUT-1 and GLUT-2 mRNA, protein, and glucose transporter activity in cultured fetal and adult hepatocytes." American Journal of Physiology-Endocrinology and Metabolism 267, no. 1 (1994): E88—E94. http://dx.doi.org/10.1152/ajpendo.1994.267.1.e88.

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To understand glycogenesis in the fetal hepatocyte, we examined glucose transport in cultured fetal and adult male rat hepatocytes. GLUT-1 mRNA was detected in fetal hepatocytes at isolation but in adult hepatocytes only after culture. GLUT-1 mRNA was more abundant in fetal than in adult hepatocytes (P < 0.005). GLUT-1 protein paralleled its message. GLUT-2 mRNA was more abundant in adult than in fetal hepatocytes (P < 0.05), and abundance did not change during culture, but GLUT-2 protein was discordantly regulated. There was more GLUT-2 protein in fetal hepatocytes at 45 h (P < 0.025

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28

Serralta, Alfonso, Maria Teresa Donato, Amparo Martinez, et al. "Influence of Preservation Solution on the Isolation and Culture of Human Hepatocytes from Liver Grafts." Cell Transplantation 14, no. 10 (2005): 837–43. http://dx.doi.org/10.3727/000000005783982495.

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A major problem for the isolation and transplantation of hepatocytes is the lack of resources for obtaining viable hepatocytes. Improving this situation would enhance hepatic cell transplantation programs. Our objective was to evaluate the influence of the preservation solutions used during organ retrieval on the quality of hepatocytes isolated from liver tissue. We compared the results of the collagenase perfusion technique for isolation of hepatocytes in human livers flushed with University of Wisconsin (UW) and Celsior preservation solutions. Yield (number of viable cells per gram of tissue

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29

Batchu, Sindhura T., Deviana Burhan, Rasika Patkar, and Bruce Wang. "Sa1497 NOVEL TECHNIQUE FOR ISOLATION OF HEPATOCYTES FROM NEEDLE BIOPSIES TO STUDY HEPATOCYTE HETEROGENEITY." Gastroenterology 158, no. 6 (2020): S—1311. http://dx.doi.org/10.1016/s0016-5085(20)33942-1.

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30

Suzuki, Atsushi, Hiromitsu Nakauchi, and Hideki Taniguchi. "In Vitro Production of Functionally Mature Hepatocytes from Prospectively Isolated Hepatic Stem Cells." Cell Transplantation 12, no. 5 (2003): 469–73. http://dx.doi.org/10.3727/000000003108747028.

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Hepatocyte transplantation and artificial organ hepatic support require a number of functionally mature hepatocytes. However, their growth activity and functional behaviors are much smaller in culture after isolation from the liver. We examined whether continuously differentiating hepatocytes from multipotent hepatic stem cells that were isolated by using flow cytometry and propagated clonally in culture could be a source of clinical application. They actually gave rise to cells that were functionally equal to mature hepatocytes found in the adult liver, which secreted albumin into culture med

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31

Fuller, Gerald M., Robert J. Bunzel, Barry M. Woloski, and Sang-Uk Nham. "Isolation of hepatocyte stimulating factor from human monocytes." Biochemical and Biophysical Research Communications 144, no. 2 (1987): 1003–9. http://dx.doi.org/10.1016/s0006-291x(87)80063-3.

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32

Dunn, Ty B., Luca Cicalese, Cristiana Rastellini, et al. "USE OF UNTRANSPLANTABLE LIVERS FOR HUMAN HEPATOCYTE ISOLATION." Transplantation 69, Supplement (2000): S180. http://dx.doi.org/10.1097/00007890-200004271-00257.

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33

Falcieri, Elisabetta, Rosalba Del Coco, Adriana R. Mariani, Pietro Gobbi, and Patrizia Santi. "Membrane modifications in the course of hepatocyte isolation." Cytotechnology 4, no. 3 (1990): 251–59. http://dx.doi.org/10.1007/bf00563785.

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34

Geerts, Sharon, Sinan Ozer, Chris Chu, et al. "Exploring donor demographics effects on hepatocyte yield and viability: Results of whole human liver isolation from one center." TECHNOLOGY 07, no. 01n02 (2019): 1–11. http://dx.doi.org/10.1142/s2339547819500018.

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Due to the growth of cell-based therapeutic alternatives addressing the shortage of livers for transplant, there is necessity for a reliable source of human hepatocytes. In addition, pharmaceutical research often requires human hepatocytes to assess new drug therapies during development or to achieve FDA approval. Whole human livers producing large quantities of cells from the same donor are ideal, enhancing reproducibility for all purposes, while also allowing for capturing variances in drug-metabolism across different demographics for pharmaceutical testing and development but are limited in

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35

Matsushita, Takakazu, Toshikazu Yagi, Joseph A. Hardin, et al. "Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver." Cell Transplantation 12, no. 2 (2003): 109–21. http://dx.doi.org/10.3727/000000003108746696.

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We have previously shown that cryopreservation leads to increased apoptotic death of porcine hepatocytes intended for use in a bioartificial liver (BAL). This study was designed to determine if a broad-spectrum caspase inhibitor, IDN–1965, reduced apoptosis and increased function of cryopreserved porcine hepatocytes in static culture or in a BAL. Porcine hepatocytes were studied immediately after isolation and after 2 weeks of cryopreservation in liquid nitrogen using medium supplemented with 25 μmol/L IDN-1965 or vehicle. Both apoptotic and necrotic cells were observed in cultures of fresh an

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36

WANG, Xiu-Jun, P. Conrad HODGKINSON, C. Matthew WRIGHT, and J. Alan PAINE. "Temperature-sensitive mRNA degradation is an early event in hepatocyte de-differentiation." Biochemical Journal 328, no. 3 (1997): 937–44. http://dx.doi.org/10.1042/bj3280937.

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The isolation and culture of metabolically active hepatocytes by proteolytic digestion of the extracellular matrix of the liver results in the transcriptional silencing of liver-specific genes encoding cytochromes P-450 (CYP) and albumin together with an induction of cellular RNase activity. The levels of albumin mRNA are maintained in cultured hepatocytes at similar levels to that present in the intact liver for at least 24 h, whereas the major constitutively expressed CYP2C11 mRNA is rapidly degraded. Hepatocytes heat-shocked at 40 °C during the isolation procedure (which results in an induc

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37

Gerlach, J. C., N. Schnoy, J. Vienken, M. Smith, and P. Neuhaus. "Comparison of Hollow Fibre Membranes for Hepatocyte Immobilisation in Bioreactors." International Journal of Artificial Organs 19, no. 10 (1996): 610–16. http://dx.doi.org/10.1177/039139889601901009.

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Various hollow fibre membranes of polyamide, cellulose and polypropylene were investigated as potential substrata for hepatocyte immobilisation in bioreactors for hybrid liver support systems. Membranes were subjected to a cytocompatibility test in which the attachment and morphology of primary hepatocytes were evaluated. The effect of coating with collagen and fibronectin was also studied. Adequate cell immobilisation was possible on polypropylene and polyamide membranes even without coating. The flattening process of the cells was dependent on the material and the coating. The incorporation

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38

Williams, Dominic P. "Application of hepatocyte-like cells to enhance hepatic safety risk assessment in drug discovery." Philosophical Transactions of the Royal Society B: Biological Sciences 373, no. 1750 (2018): 20170228. http://dx.doi.org/10.1098/rstb.2017.0228.

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Hepatic stress and injury from drugs continues to be a major concern within the pharmaceutical industry, leading to preclinical and clinical attrition precautionary warnings and post-market withdrawal of drugs. There is a requirement for more predictive and mechanistically accurate models to aid risk assessment. Primary human hepatocytes, subject to isolation stress, cryopreservation, donor-to-donor variation and a relatively short period of functional capability in two-dimensional cultures, are not suitable for high-throughput screening procedures. There are two areas within the drug discover

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39

G., Durga Prameela, T. N. V. K. V. Prasad*, M. Nagalakshmi Devamma, E. K. Elumalai, P. M. Ayyasamy, and T. Kiran Reddy. "Isolation and characterization of hematopoietic stem cells from human umbilical cord blood for cell therapy." International Journal of Bioassays 1, no. 10 (2012): 107. http://dx.doi.org/10.21746/ijbio.2012.10.0010.

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Alternatives to organ transplantation for treatment of certain liver diseases are currently under investigation. Whole liver, split liver and related living donor liver transplantation are clinically safe and well established procedures for the treatment of end-stage liver failure. However, organ donor shortage and the need of lifelong immunosuppressive treatment are still the major limitations of all these options. Therefore, the development of cell based therapeutic strategies for liver disease is under experimental evaluation. Umbilical cord blood was collected and isolated the mononucleate

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40

Schön, Michael R., Gero Puhl, Jörg Gerlach, Jorn Frank, and Peter Neuhaus. "Hepatocyte isolation from pig livers after warm ischaemic injury." Transplant International 7, s1 (1994): 159–62. http://dx.doi.org/10.1111/j.1432-2277.1994.tb01337.x.

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41

Izamis, Maria-Louisa, Sinem Perk, Candice Calhoun, Korkut Uygun, Martin L. Yarmush, and François Berthiaume. "Machine perfusion enhances hepatocyte isolation yields from ischemic livers." Cryobiology 71, no. 2 (2015): 244–55. http://dx.doi.org/10.1016/j.cryobiol.2015.07.006.

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42

Figueiredo, Neusa, Beatriz Matos, Mário Diniz, Vasco Branco, and Marta Martins. "Marine Fish Primary Hepatocyte Isolation and Culture: New Insights to Enzymatic Dissociation Pancreatin Digestion." International Journal of Environmental Research and Public Health 18, no. 4 (2021): 1380. http://dx.doi.org/10.3390/ijerph18041380.

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Primary cell cultures from wild organisms have been gaining relevance in ecotoxicology as they are considered more sensitive than immortalized cell lines and retain the biochemical pathways found in vivo. In this study, the efficacy of two methods for primary hepatocyte cell isolation was compared using liver from two marine fish (Sparus aurata and Psetta maxima): (i) two-step collagenase perfusion and (ii) pancreatin digestion with modifications. Cell cultures were incubated in L-15 medium at 17 ± 1 °C and monitored for up to six days for cell viability and function using the trypan blue excl

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Belaschk, Elisa, Susanne Rohn, Ramla Mukiibi, et al. "Isolation, Characterization and Cold Storage of Cells Isolated from Diseased Explanted Livers." International Journal of Artificial Organs 40, no. 6 (2017): 294–306. http://dx.doi.org/10.5301/ijao.5000594.

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Introduction Livers discarded after standard organ retrieval are commonly used as a cell source for hepatocyte transplantation. Due to the scarcity of organ donors, this leads to a shortage of suitable cells for transplantation. Here, the isolation of liver cells from diseased livers removed during liver transplantation is studied and compared to the isolation of cells from liver specimens obtained during partial liver resection. Methods Hepatocytes from 20 diseased explanted livers (Ex-group) were isolated, cultured and stored at 4°C for up to 48 hours, and compared to hepatocytes isolated fr

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Gerlach, J. C., J. Brombacher, J. M. Courtney, and P. Neuhaus. "Nonenzymatic versus Enzymatic Hepatocyte Isolation from Pig Livers for Larger Scale Investigations of Liver Cell Perfusion Systems." International Journal of Artificial Organs 16, no. 9 (1993): 677–81. http://dx.doi.org/10.1177/039139889301600909.

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A comparison of nonenzymatic and enzymatic hepatocyte isolation was performed on pig livers. The collagenase perfusion showed superior results: mean viability 72 ± 10% versus a maximum viability of 21% using EDTA-perfusion. A five-step collagenase perfusion technique, developed for pig livers enables larger scale investigations, in order to develop methods for hepatocyte cultures in therapeutical liver cell perfusion systems.

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Stamatoglou, S. C., R. C. Hughes, and U. Lindahl. "Rat hepatocytes in serum-free primary culture elaborate an extensive extracellular matrix containing fibrin and fibronectin." Journal of Cell Biology 105, no. 5 (1987): 2417–25. http://dx.doi.org/10.1083/jcb.105.5.2417.

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Adult rat hepatocytes cultured on type IV collagen, fibronectin, or laminin and maintained in serum-free medium were examined by indirect immunofluorescence using polyclonal antibodies against extracellular matrix proteins. An extensive fibrillar matrix containing fibronectin and fibrin was detected in all hepatocyte cultures irrespective of the exogenous matrix substratum used to support cell adhesion. Fibrils radiated from the cell periphery and covered the entire culture substratum. In addition, thicker fibers or bundles of fibers were localized on top of hepatocytes. This matrix did not co

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Kumashiro, Y., K. Teramoto, K. Shimizu-Saito, K. Asahina, H. Teraoka, and S. Arii. "Isolation of hepatocyte-like cells from mouse embryoid body cells." Transplantation Proceedings 37, no. 1 (2005): 299–300. http://dx.doi.org/10.1016/j.transproceed.2005.01.036.

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Kluge, Martin, Anja Reutzel-Selke, Hendrik Napierala, et al. "Human Hepatocyte Isolation: Does Portal Vein Embolization Affect the Outcome?" Tissue Engineering Part C: Methods 22, no. 1 (2016): 38–48. http://dx.doi.org/10.1089/ten.tec.2015.0190.

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Kumashiro, Yuji, Kenichi Teramoto, Keiko Shimizu-Saito, Kinji Asahina, Hirobumi Teraoka, and Shigeki Arii. "ISOLATION OF HEPATOCYTE-LIKE CELLS FROM MOUSE EMBRYOID BODY CELLS." Transplantation 78 (July 2004): 594. http://dx.doi.org/10.1097/00007890-200407271-01599.

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Riou, J. P., C. Audigier, M. Laville, M. Beylot, P. Pigeon, and R. Mornex. "Dephosphorylation of l-pyruvate kinase during rat liver hepatocyte isolation." Archives of Biochemistry and Biophysics 236, no. 1 (1985): 321–27. http://dx.doi.org/10.1016/0003-9861(85)90632-0.

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Gerlach, J. C. "Development of a Hybrid Liver Support System: A Review." International Journal of Artificial Organs 19, no. 11 (1996): 645–54. http://dx.doi.org/10.1177/039139889601901105.

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Hybrid liver support systems (LSS) for the use of the detoxifying, metabolic synthetic and regulatory capabilities of liver cells are under development for extracorporeal therapy of acute liver failure and for bridging to liver transplantation. A summary of our development is discussed. A five-step technique for primary liver cell isolation has been introduced in order to address larger scale procurement of hepatocytes. Immobilisation of the cells after isolation appears to be one of the main factors in maintaining hepatocyte function in vitro. Different techniques have been investigated. Usin

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