SV40T介导的人胎儿永生化气管成纤维细胞系的建立

建立永生化气管成纤维细胞系为研究细胞恶性转化提供实验模型。用ｐＺＩＰＳＶ４０Ｔ转染原代人胎气管成纤维细胞,Ｇ４１８筛选,挑取阳性克隆传代培养,并进行细胞生物学特性鉴定。结果表明,细胞经ｐＺＩＰＳＶ４０Ｔ转染后,形态较正常细胞略短,排列不规则,当传到５０代时仍生长良好。Ｓｏｕｔｈｅｒｎ ｂｌｏｔ 证实此转化细胞整合了ＳＶ４０Ｔ基因。永生化细胞接种的第４天细胞为指数生长期,染色体核型以非整倍体为主,不

ESTABLISHMENT OF HUMAN FETAL TRACHEAL FIBROBLAST CELL LINE IMMORTALIZED BY SV40T

The aim is immortalizing the human fetal tracheal fibroblast cells to study the cell neoplastic transformation. The primary human fetal tracheal fibroblast cell was transfected with pZIPSV40T vector. The colonies that resistant to G418 were collected, and passaged to immortalize. The biological characters immortalized cell had been determined. Result: the immortalized cell morphology is shorter than normal, arrange un regular. The cells grow well at p50. Southern blot confirmed the presence of T antigen as integrated form in immortalized cell DNA. The index of growth period was fourth day after plated. All the cells could not form colony in soft agar. No tumors formed after SC injection of the cells in nude mice. The result confirmed that human fetal tracheal fibroblast cell can be immortalized by SV40T gene.