子宫内膜基质细胞作为饲养层培养人胚原始生殖细胞

A CULTURE SYSTEM USING HUMAN ENDOMETRIAL FIBROBLASTS AS FEEDER CELL ALLOWS PRODUCTION OF HUMAN PRIMODIAL GERM CELLS

目的探讨子宫内膜基质细胞作为饲养细胞培养人胚原始生殖细胞的可能。方法从5～9周人胚胎生殖嵴、中肾嵴、肠系膜中消化分离原始生殖细胞(PGCs),种植于经丝裂霉素C处理的人子宫内膜基质细胞饲养层上培养。结果PGCs可以在体外培养3个多月,传14代。这样培养的PGCs表达胚胎干细胞的多种标志,如碱性磷酸酶染色、SSEA和TRA抗原等。另外细胞保持正常的染色体核型。结论人子宫内膜基质细胞可以作为饲养细胞在体外培养人胚原始生殖细胞,保持未分化状态。

Objective To investigate whether endometrial fibroblasts as feeder cell can be used for cultivation of human primordial germ cells(PGCs). Methods Human primodial germ cells(PGCs) were isolated from the gonadal ridges and mesenteries 5-9 weeks after fertilization and were planted on to mitotically inactivated human endometrial fibroblasts. Results PGCs have been cultured more than 3 months so far for 14 passages on human endometrial fibroblast feeder layer and still positive for plutipotent marker of embryonic stem cells such as alkaline phosphates(AKP), stage-spesific embryonic antigens(SSEA) and tumor rejection antigens(TRA).These cells were rather stable in vitro and showed normal karyotype up to 14 passages.Conclusion We successfully used human endometrial fibroblasts as feeder cell for derivation and continued undifferentiated growth of PGCs.;