[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.


CONCLUSIONS
FACS can be used to isolate quantities of viable spermatogonial stem cells.
Liu
Chun-xia
CX
Department of Urology, Renmin Hospital, Wuhan University, Wuhan, Hubei 430060, China.
Zhang
Ci
C
Wang
Ling-long
LL
Yu
Xiao-xiang
XX
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D003456:Cryptorchidism; D004195:Disease Models, Animal; D005434:Flow Cytometry; D005453:Fluorescence; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D011897:Random Allocation; D013093:Spermatogonia / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*