Sorting of spermatogonial stem cells in mice / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To search for an effective method for enriching spermatogonial stem cells in mice.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>The sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable.</p><p><b>CONCLUSION</b>FACS can be used to isolate quantities of viable spermatogonial stem cells.</p>