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  • Title: [The feeder layer of human embryonic fibroblasts supports the growth of human spermatogonial stem cells].
    Author: Wang YB, Chen B, Wang YC, Zhang ZL, Wang HX, Lu YN, Xiang ZQ, Hu K, Yang YK, Han YF, Wang Z, Wang YX, Huang YR.
    Journal: Zhonghua Nan Ke Xue; 2008 Dec; 14(12):1063-8. PubMed ID: 19157223.
    Abstract:
    OBJECTIVE: To investigate the methods and conditions for the isolation, purification and culture of human spermatogonial stem cells (SSCs) on the feeder layer cells of human embryonic fibroblasts (hEFs). METHODS: SSCs isolated and purified from normal human fetal testicular tissues by sequential two-step enzyme digestion and Percoll uncontinuous density gradient centrifugation were cultured on the feeder layer cells of hEFs isolated from 5-9 weeks old human embryos. The surface markers SSEA-1 and OCT4 of the SSCs were detected by immunohistochemistry; the alkaline phosphatase (AKP) activity of the SSC clones measured; and the expressions of the SSC-related genes determined by RT-PCR. RESULTS: SSCs survived, proliferated and formed colonies on the feeder layers, and the colonies were highly positive for SSEA-1 and OCT4, with strong AKP activity and high expressions of the SSC-related genes. CONCLUSION: The feeder layer of hEFs supports the growth of human spermatogonial stem cells.
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