[Sorting of spermatogonial stem cells in mice].

OBJECTIVE

To search for an effective method for enriching spermatogonial stem cells in mice.

METHODS

Bilateral artificial cryptorchidism was performed on 20 six-week old male Kunming mice. Three months after the operation, the testes were removed and single cell suspension prepared by two-step enzyme digestion. FITC-conjugated anti-alpha6-integrin antibody and PE-conjugated anti-c-kit antibody were added for adequate time on ice. Then the cells with low side scatter light-scattering properties were sorted and positively stained for alpha6-integrin and negative c-kit expression. And the viability of the isolated cells was assessed by trypan blue exclusion.

RESULTS

The sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable.

CONCLUSION

FACS can be used to isolate quantities of viable spermatogonial stem cells.