[Establishment and mechanistic characterization of SV40 T antigen immortalized human fetal hepatocytes].
Human fetal liver cells were cultured in serum free media enriched with hydrocortisone and infected with retrovirus containing SV40 large T gene. Replicating colonies with G-418 resistance developed in 50 days through 23 passages, while none grew in control dishes. After a crisis of 3 months duration, three colonies resumed active proliferation for over 15 months through 60 passages and became immortalized. These cells had epithelioid morphology, and were stained positively for CK-18. Southern blot and immunocytochemistry demonstrated the presence and expression of SV40 T-antigen in the immortalized cell lines. The cells expressed human albumin, especially in those while in cycle. Accumulation of p53 protein in the cell nuclei and strong expression of TGF-alpha as shown by immunocytochemistry explained at least partly the mechanism of unlimited growth of these immortalized human hepatocytes. These cells did not show anchorage-dependent growth in soft agar, nor did tumor form when inoculated into nude mice. These immortalized, differentiated, non-malignant human fetal hepatocyte lines may be useful for further studies.
Wu
G S
GS
Cancer Institute, CAMS & PUMC, Beijing.
D000418:Albumins / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D005333:Fetus; D015870:Gene Expression; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D012190:Retroviridae / Q000276:immunology*; D013539:Simian virus 40 / Q000276:immunology; D014162:Transfection; D016211:Transforming Growth Factor alpha / Q000096:biosynthesis / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics
D000418:Albumins; D000952:Antigens, Polyomavirus Transforming; D016211:Transforming Growth Factor alpha; D016159:Tumor Suppressor Protein p53