Optimization of Conditions for Clinical Human Hepatocyte Infusion
| العنوان: | Optimization of Conditions for Clinical Human Hepatocyte Infusion |
|---|---|
| المؤلفون: | Dawen Bu, Robert A. Fisher, Melissa T. Thompson, Luke G. Wolfe, Joseph K. Ritter |
| المصدر: | Cell Transplantation, Vol 13 (2004) |
| بيانات النشر: | SAGE Publishing, 2004. |
| سنة النشر: | 2004 |
| مصطلحات موضوعية: | 0301 basic medicine, Male, Adenosine, Adolescent, Cell Survival, Cell Transplantation, Biomedical Engineering, Cell Culture Techniques, Gene Expression, lcsh:Medicine, Apoptosis, DNA Fragmentation, Culture Media, Serum-Free, Andrology, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Cytochrome P-450 Enzyme System, medicine, Cytochrome P-450 CYP3A, Humans, MTT assay, Viability assay, Aspartate Aminotransferases, Glucuronosyltransferase, Child, Cell Proliferation, Transplantation, L-Lactate Dehydrogenase, Endoplasmic reticulum, Fatty liver, lcsh:R, Cell Biology, Middle Aged, medicine.disease, Fatty Liver, 030104 developmental biology, medicine.anatomical_structure, chemistry, Hepatocyte, Hepatocytes, Trypan blue, Female, 030217 neurology & neurosurgery |
| الوصف: | Cytotoxicity and apoptosis are common problems in the isolation and storage of human hepatocytes. In vitro environments of hepatocytes during cell infusion may be critical to reducing cellular damage and enhancing cell viability. We examined the effects of donor liver histology (40–50% steatosis vs. normal), incubation time, temperature, and three solutions for infusion on banked primary human hepatocytes, by studying: trypan blue exclusion, AST release, LDH release, MTT assay, detection of DNA ladder, and a hepatocyte proliferation assay. In addition, the microstructure functions of the endoplasmic reticulum and mitochondria of the intact hepatocytes were determined by measuring correlates of UGT 1A1 and cytochrome P-450 3A (CYP3A4) activity. In general, hepatocyte viability decreased significantly within 60 min after thawing. Cells suspended in 5% dextrose lactated Ringers solution (D5LR) maintained greater cell viability. Hepatocytes from normal liver donors showed less AST and LDH enzyme leak in comparison with cells from fatty liver donors. Mild hypothermic temperature (32°C) inhibited cellular damage that otherwise significantly increased at 60 min. Hepatocytes did not proliferate until 12 h from thaw, regardless of supernatant or conditions of suspension. CYP3A4 activity and a marker for UGT 1A1 activity in hepatocytes from normal donor livers were higher than those from steatotic donor livers. These findings suggest that hepatocytes suspended for infusion after isolation from normal liver donors have normal biological functions and less cellular damage/necrosis in contrast with those isolated from fatty liver donors. These damages are inhibited significantly by maintaining hepatocytes at a mild hypothermic temperature (32°C). D5LR alone maintained the best cell viability for up to 60 min. Media of D5LR + adenosine and HMM were able to partially inhibit hepatocyte apoptosis in hepatocytes from steatotic livers. |
| اللغة: | English |
| تدمد: | 1555-3892 0963-6897 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::880a462188bff1f1f8b15bb33e7f715b https://doaj.org/article/2e3c70aa587a4b7581fdd34ebf841164 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....880a462188bff1f1f8b15bb33e7f715b |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15553892 09636897 |
|---|