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Long-term proliferation and characterization of human spermatogonial stem cells obtained from obstructive and non-obstructive azoospermia under exogenous feeder-free culture conditions


D. R. Lee, PhD, Fertility Center, CHA Gangnam Medical Center, CHA University, 606-5 Yeoksam-dong, Gangnam-gu, Seoul 135-081, Korea. Tel.: +82-2-3468-3421; Fax: +82-2-3468-2610; E-mail:

K.-S. Kim, DVM, PhD, Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Seoul 133–791, Korea. Tel.: +82-2-2220-0607; Fax: +82-2-2281-7841; E-mail:


Objectives:  The aim of the present study was to improve efficiency of isolation and to optimize proliferative potential of human spermatogonial stem cells (SSCs) obtained from obstructive azoospermic (OA) and non-obstructive azoospermic (NOA) patients, and further, to characterize these cells for potential use in infertility treatment or study of reproductive biology.

Materials and methods:  We have applied a cell-sorting method, using collagen and magnetic activated cell separation to overcome obstacles, developing a collection system, and simple long-term proliferation system, that yields large numbers of high-purity SSCs from obstructive OA and NOA patients.

Results:  SSCs derived from OA and NOA patients proliferated and maintained their characteristics for more than 12 passages (>6 months) in vitro. Moreover, the population of cells positive for the SSC-specific markers GFRα-1 and integrin α6, increased to more than 80% at passage 8.

Conclusion:  These finding may support the idea that in vitro propagation of SSCs could be a useful tool for infertility treatment and study of reproductive biology.