The feeder layer of human embryonic fibroblasts supports the growth of human spermatogonial stem cells / 中华男科学杂志
National Journal of Andrology
;
(12): 1063-1068, 2008.
Article
in Chinese
| WPRIM
| ID: wpr-309738
ABSTRACT
<p><b>OBJECTIVE</b>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).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>The feeder layer of hEFs supports the growth of human spermatogonial stem cells.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Spermatogonia
/
Stem Cells
/
Cell Differentiation
/
Cells, Cultured
/
Cell Culture Techniques
/
Cell Biology
/
Embryo, Mammalian
/
Fibroblasts
/
Methods
Limits:
Humans
/
Male
Language:
Chinese
Journal:
National Journal of Andrology
Year:
2008
Type:
Article
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