Wu G S
Cancer Institute, CAMS & PUMC, Beijing.
Zhonghua Zhong Liu Za Zhi. 1993 Nov;15(6):415-8.
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.
将人胎肝细胞培养于富含氢化可的松的无血清培养基中，并用含有SV40大T基因的逆转录病毒感染。在50天内经过23代培养，出现了对G - 418有抗性的复制菌落，而对照培养皿中没有生长。经过3个月的危机期后，三个菌落恢复活跃增殖，经过60代培养超过15个月并永生化。这些细胞具有上皮样形态，CK - 18染色呈阳性。Southern印迹和免疫细胞化学证明永生化细胞系中存在SV40 T抗原并表达。这些细胞表达人白蛋白，尤其是处于细胞周期中的细胞。免疫细胞化学显示细胞核中p53蛋白的积累和TGF -α的强表达至少部分解释了这些永生化人肝细胞无限生长的机制。这些细胞在软琼脂中不显示锚定依赖性生长，接种到裸鼠中也不形成肿瘤。这些永生化、分化的非恶性人胎肝细胞系可能有助于进一步研究。
