Abstract:
: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 leakage of lactate dehydrogenase (LD) and aspartate aminotransferase (AST), lidocaine clearance, albumin production, and flow cytometry. With the centrifuge method the mean cell viability was 92.5%, while with the BRAT method the viability was 95.9%. The minimal cell yields with the BRAT procedure were 7.3 x 10(9) for 250-ml centrifuge bowls and 2.8 x 10(9) for 165-ml bowls, which compares well with that found by other authors. Because the same initial procedures were employed in both methods the total hepatocyte yield per liver was comparable. Flow cytometry confirmed that the proliferation of hepatocytes was facilitated by oxygenation during the isolation procedure. The recovery of hepatocytes in culture following isolation was similar after either method. Daily microscopic investigation indicated that cytoplasmic vacuolization and granularities were present after either procedure and these disappeared following 3-4 days of culturing. Flow cytometry indicated that the hepatocyte cell cycle was similar after either method; at 7 days the profile indicated that the cells were still proliferating. Trends in the lactate to pyruvate ratio and the leakage of LD and AST indicated that the functional polarity of hepatocytes was regained after approximately 3 days. Lidocaine clearance at 4 days indicated that the cytochrome P450 system was active, while significant albumin production was apparent at day 5. The benefit of using BRAT technology in hepatocyte isolation lies in guaranteed sterility, convenience, speed, and the ability to oxygenate media and cell suspensions during the procedure.
journal_name
Cell Transplantjournal_title
Cell transplantationauthors
Nieuwoudt MJ,Kreft E,Olivier B,Malfeld S,Vosloo J,Stegman F,Kunneke R,Van Wyk AJ,Van der Merwe SWdoi
10.3727/000000005783983007subject
Has Abstractpub_date
2005-01-01 00:00:00pages
291-9issue
5eissn
0963-6897issn
1555-3892journal_volume
14pub_type
杂志文章abstract::Cell and tissue culture techniques have improved considerably since the first attempts to maintain explants of animal tissue in vitro. The two major developments that have allowed these improvements are the ability to produce continuous cell lines, thus allowing reproducible results to be obtained, and the definition ...
journal_title:Cell transplantation
pub_type: 杂志文章,评审
doi:10.1016/0963-6897(95)02005-5
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abstract::Stroke is one of the most devastating diseases worldwide. In recent years, a great number of studies have focused on the effects of microRNAs (miRNAs) on stroke and the results demonstrated that the expressions of miRNAs are associated with the prognosis of stroke. In the present study, we review relevant articles reg...
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abstract::Multilineage-differentiating stress-enduring (Muse) cells are endogenous pluripotent stem cells that can be isolated based on stage-specific embryonic antigen-3 (SSEA-3), a pluripotent stem cell-surface marker. However, their capacities for survival, neurotrophic factor secretion, and neuronal and glial differentiatio...
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doi:10.3727/096368916X690421
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abstract::Several types of hair loss result from the inability of hair follicles to initiate the anagen phase of the hair regeneration cycle. Modulating signaling pathways in the hair follicle niche can stimulate entry into the anagen phase. Despite much effort, stem cell-based or pharmacological therapies to activate the hair ...
journal_title:Cell transplantation
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pub_type: 杂志文章,已发布勘误
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更新日期:2016-07-01 00:00:00
abstract::The use of regenerative medicine to treat nervous system disorders like ataxia has been proposed to either replace or support degenerating neurons. In this study, we assessed the ability of human neural progenitor cells (hNPCs) to repair and restore the function of dying neurons within the spastic Han-Wistar rat (sHW)...
journal_title:Cell transplantation
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doi:10.1177/0963689717723637
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更新日期:1996-09-01 00:00:00
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journal_title:Cell transplantation
pub_type: 杂志文章
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doi:10.3727/096368916X691682
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journal_title:Cell transplantation
pub_type: 杂志文章
doi:10.1016/0963-6897(96)00026-7
更新日期:1996-07-01 00:00:00
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journal_title:Cell transplantation
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abstract::To overcome common disadvantages of standard cultured epidermal sheet grafts (CEG) we have developed a new technique of transplanting cultured human keratinocytes suspended as single cells in a fibrin-glue matrix (Keratinocyte-fibrin-glue suspension-KFGS). In an athymic mouse model with reproducible standardized full ...
journal_title:Cell transplantation
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更新日期:1998-05-01 00:00:00
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journal_title:Cell transplantation
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doi:
更新日期:2007-01-01 00:00:00
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doi:10.3727/096368911X637443
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journal_title:Cell transplantation
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journal_title:Cell transplantation
pub_type: 杂志文章
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更新日期:2001-01-01 00:00:00
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journal_title:Cell transplantation
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journal_title:Cell transplantation
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pub_type: 杂志文章
doi:10.1177/096368970000900101
更新日期:2000-01-01 00:00:00
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journal_title:Cell transplantation
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doi:10.3727/000000008783906928
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abstract::Testis-derived Sertoli cells have been used to create an immune "privileged" site outside of the testis to facilitate cell transplantation protocols for diabetes and neurodegenerative diseases. In addition to secreting immunoprotective factors, Sertoli cells also secrete growth and trophic factors that appear to enhan...
journal_title:Cell transplantation
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