SNL细胞作为饲养层细胞培养人胚胎干细胞

SNL cells as new feeder cells to culture human embryonic cells

目的为便于人胚胎干细胞(ES细胞)的培养和研究,寻找一种建系细胞作为人ES细胞的饲养层细胞。方法以小鼠成纤维细胞系SNL细胞作为饲养层细胞培养人ES细胞株hES1。传代17代后,检测hES细胞表面特异性标志,包括碱性磷酸酶(AKP)、Oct-4、阶段特异性胚胎抗原(SSEA)-3和SSEA-4的表达;同时观察细胞的多向分化潜能。结果hES1细胞在SNL细胞饲养层上能持续生长。经多次传代后持续表达人ES细胞表面特异性标志,其注射于重症联合免疫缺陷小鼠皮下仍能形成畸胎瘤组织。结论SNL细胞能够作为饲养层细胞用于人ES细胞的培养,且操作更方便;该细胞已转染了耐受新霉素的基因,为将来对人ES细胞基因操作时进行药物筛选提供了可能。

Objective To look for a new feeder cells to facilitate culturing the human embryonic stem cells（ES cells）. Methods SNL cells （ permanent line of irradiated mouse fibroblast cells） were tested as feeder cells to culture the human ES cells. After growing on SNL cells for 17 passages, the human ES cells were tested for the expressing of the markers of stem cells such as alkaline phosphatase （AKP） , Oct-4, stage-specific embryonic antigen （SSEA）-3, and SSEA-4. The ES cells were injected into SCID mice to test their pluripotent ability. Results Human ES cells grew well on feeder cells of SNL. The clones growing on SNL cells were thinner than those on normal MEF cells. The outcomes of staining showed that after 17 passages, the human ES cells continued to express AKP, Oct-4, SSEA- 4 and SSEA-3. When injected subcutaneously into SCID mice the human ES cells developed into the teratoma. Conclusion SNL cells can be used as feeder cells to culture human ES cells. The SNL cells are more convenient than normal MEF cells as the feeder cells to culture human ES cells, and can be gene modified to resist the G418 so as to facilitate gene modifying study of human ES cells.