人睾丸支持细胞的分离培养及鉴定

Isolation,Culture and Identification of Human Sertoli Cells

目的:建立人胚胎睾丸支持细胞体外分离,纯化及培养的方法,并进行鉴定。方法:取胎龄13~28周的胚胎睾丸,采用两步酶消化,添加特定培养基,培养过程中纯化睾丸支持细胞,并用免疫荧光及RT-PCR技术对培养细胞进行鉴定。结果:通过检测阶段特异性标记物M IS提示胚胎睾丸支持细胞占培养细胞总数的90%以上,经RT-PCR及免疫荧光检测所培养细胞具有分泌产生SCF的能力。结论:在本研究建立的培养体系下细胞可稳定传代,经鉴定为高纯度的睾丸支持细胞,并维持了其体内生物学活性。

Objective The aim of our study was to establish a method of isolation,purification culture and identification for human fetal Sertoli cells in vitro.Methods The testes were obtained from fetuses of 13～28 week of gestation.Testis cells were collected by two-step enzymatic digestion and cultivated with a modified system.The Sertoli cells were purified in the culture system,and immunofluorescence staining and RT-PCR assay were carried for the identification of the cultured cells.Results The purity of the cultu...