Enrichment of type A1-A4 spermatogonia by flow cytometry.
- Author:
Yang BAI
1
;
Zhe-Wei YE
;
Fu-Qing ZENG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Separation; methods; Flow Cytometry; methods; Male; Microscopy, Electron; Proto-Oncogene Proteins c-kit; biosynthesis; Rats; Rats, Sprague-Dawley; Spermatogonia; cytology; metabolism; ultrastructure; Stem Cells; cytology; metabolism; ultrastructure; Testis; cytology; metabolism
- From: National Journal of Andrology 2008;14(1):3-6
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore a method to isolate and purify the subtype of type A spermatogonial stem cells (SSCs).
METHODSWe 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.
RESULTSThe 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.
CONCLUSIONWith c-kit as the marker, FACS can effectively isolate and purify the subtype of SSCs after preliminarily purified by discontinuous density gradient centrifugation.
