Immortalization of multipotent stem cells by using the oncogene transgenic mice

为建立多种永生化多能干细胞系，利用携带猴病毒40（SV 40）温度敏感性T抗原（TS抗原）基因的转基因小鼠，将分选后的干细胞与骨髓基质细胞共培养，建立多能造血干细胞系。一种细胞系是B淋巴干细胞系，并且自我更新以及定向分化为更成熟的B细胞。其他细胞系来自转基因小鼠，具有干细胞的表面标记，并且可以分化为更成熟的造血细胞。制备了几种带有bcl-2基因、c-kit基因或Sl基因的转基因小鼠，并用于永生化原始生殖细胞，并且来自这些转基因小鼠的生殖细胞显示出它们在培养中的长期维持，转基因小鼠的胃粘膜细胞也具有正常胃粘膜细胞的功能，包括产生保护性因子，如糖蛋白、碳酸氢盐和胰头素。

To establish a variety of immortalized multipotent stem cell lines, the transgenic mice harboring temperature-sensitive T-antigen (ts-antigen) gene of simian virus 40 (SV40) were used.To establish multipotent hematopoietic stem cell lines, the sorted stem cells were cocultured with the bone marrow stromal cell lines and two immature hematopoietic stem-like cell lines were immortalized depending on thr stromal cells. One cell line is a B-lymphoid stem cell line and self-renewed as well as committed to differentiate more mature B cells. The other cell lines were obtained from the transgenic mice and exhibited surface markers of the stem cells and can be differentiated to more mature hematopoietic cells.Several transgenic mice with bcl-2 gene, c-kit gene, or Sl gene were made and used to immortalize primordial germ cells, and the germ cells from such transgenic mice showed their prolonged maintenance in culture, but the immortalization was not yet observed.Gastric mucosal cells were also established fro the transgenic mice and showed specific functions of normal gastric mucosal cells including production of protective factors such as glycoproteins, bicarbonate, and prostaglandins.