Enrichment of type A1-A4 spermatogonia by flow cytometry / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To explore a method to isolate and purify the subtype of type A spermatogonial stem cells (SSCs).</p><p><b>METHODS</b>We isolated spermatogonia by discontinuous density gradient centrifugation, sorted c-kit-expressed cells with the fluorescence-activated cell sorter (FACS), observed their ultrastructure by electron microscope, and performed immunohistochemistry to determine the expression of c-kit in the testis.</p><p><b>RESULTS</b>The c-kit positive cells constituted (18.65 +/- 1.69) % of the testis cells that were isolated by density gradient centrifugation, but made up only (3.16 +/- 0.84) % of those that were not (P < 0.01). The rates of recovery and viability of the c-kit positive cells sorted by FACS were (65.90 +/- 1.24)% and (85.60 +/- 1.14)%, respectively.</p><p><b>CONCLUSION</b>With c-kit as the marker, FACS can effectively isolate and purify the subtype of SSCs after preliminarily purified by discontinuous density gradient centrifugation.</p>