RRC ID 12032
Author Hoekstra R, Deurholt T, ten Bloemendaal L, Desille M, van Wijk AC, Clement B, Oude Elferink RP, van Gulik TM, Chamuleaut RA.
Title Assessment of in vitro applicability of reversibly immortalized NKNT-3 cells and clonal derivatives.
Journal Cell Transplant
Abstract In vitro applications of human hepatocytes, such as bioartificial livers and toxicity assays, require thoroughly testing of human cell lines prior to using them as alternative cell sources. The reversibly immortalized NKNT-3 cell line was reported to show clear in vivo functionality. Here, NKNT-3 cells were tested for their in vitro applicability. Low-passage (P2) and high-passage (P28) NKNT-3 cells and clonal derivatives were characterized for reversion of immortalization, heterogeneity, and hepatic functionality. Reversion with reduced expression of immortalizing agent could be established. However, during culturing the cells lost the capacity to be selected for completed reversion. The phenotypic instability is probably associated with heterogeneity in the culture, as clonal derivatives of P2 cells varied in morphology, growth, and reversion characteristics. The mRNA levels of genes related with hepatic differentiation increased 4-20-fold after reversion. However, the levels never exceeded 0.1% of that detected in liver and no urea production nor ammonia elimination was detected. Additionally, activities of different cytochrome P450s were limited. In conclusion, the NKNT-3 culture is heterogeneous and unstable and the in vitro functionality is relatively low. These findings emphasize that in vivo testing of hepatic cell lines is little informative for predicting their value for in vitro applications.
Volume 15(5)
Pages 423-33
Published 2006-1-1
PMID 16970284
MeSH Cell Culture Techniques / methods Cell Differentiation / genetics Cell Differentiation / physiology Cell Line Clone Cells / cytology Clone Cells / metabolism Gene Expression / genetics Glutathione Transferase / genetics Glutathione Transferase / metabolism Hepatocytes / cytology* Hepatocytes / metabolism* Hepatocytes / transplantation Humans Liver / cytology Liver / metabolism Proteins / metabolism RNA, Messenger / genetics RNA, Messenger / metabolism Reverse Transcriptase Polymerase Chain Reaction Time Factors alpha-Fetoproteins / genetics alpha-Fetoproteins / metabolism
IF 3.341
DNA material AxCANCre (RDB01748)