Sorting of spermatogonial stem cells in mice.
- Author:
Chun-xia LIU
1
;
Ci ZHANG
;
Ling-long WANG
;
Xiao-xiang YU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Separation; methods; Cryptorchidism; Disease Models, Animal; Flow Cytometry; Fluorescence; Male; Mice; Mice, Inbred Strains; Random Allocation; Spermatogonia; cytology; Stem Cells; cytology
- From: National Journal of Andrology 2007;13(1):24-26
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo search for an effective method for enriching spermatogonial stem cells in mice.
METHODSBilateral 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.
RESULTSThe sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable.
CONCLUSIONFACS can be used to isolate quantities of viable spermatogonial stem cells.
