Document Detail

In vitro culture and morphological characterization of prepubertal buffalo (Bubalus bubalis) putative spermatogonial stem cell.
MedLine Citation:
PMID:  23151879     Owner:  NLM     Status:  MEDLINE    
PURPOSE: Spermatogonial stem cells (SSCs) have the unique ability both to self-renew and to produce progeny that undergo differentiation to spermatozoa. The present study has been carried out to develop a method to purify and enrich the pure populations of spermatogonial stem cell like cells in buffalo.
METHODS: The spermatogonial cells were isolated from testes of 3-7 month old buffalo calves and disaggregated by double enzymatic digestion. Mixed population of isolated cells were then plated on Datura stramonium agglutinin (DSA) lectin coated dishes for attachment of Sertoli cells. The desired cells were obtained from suspension medium after 18 h of incubation and then loaded on discontinuous density gradient using percoll (20-65 %) and different types of spermatogonia cells were obtained at interface of each layer. These cells were cultured in vitro.
RESULTS: Spermatogonial cells isolated have spherical outline and two or three eccentrically placed nucleoli, created a colony after proliferation during first week or immediately after passage. After 7-10 days of culture, the resulted developed colonies of spermatogonial cells expressed the spermatogonial specific genes like Plzf and VASA; and other pluripotency related markers viz. alkaline phosphtase, DBA, CD9, CD90, SSEA-1, OCT-4, NANOG and REX-1.
CONCLUSION: Our results show that the isolated putative spermatogonial stem cells exhibit the expression of pluripotency related and spermatogonial specific genes. This study may help to establish a long term culture system for buffalo spermatogonia.
S Kala; R Kaushik; K P Singh; P H Kadam; M K Singh; R S Manik; S K Singla; P Palta; M S Chauhan
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-11-15
Journal Detail:
Title:  Journal of assisted reproduction and genetics     Volume:  29     ISSN:  1573-7330     ISO Abbreviation:  J. Assist. Reprod. Genet.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-24     Completed Date:  2013-07-12     Revised Date:  2013-12-04    
Medline Journal Info:
Nlm Unique ID:  9206495     Medline TA:  J Assist Reprod Genet     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1335-42     Citation Subset:  IM    
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MeSH Terms
Cell Culture Techniques*
Cell Differentiation
Cell Lineage*
Cells, Cultured
Sertoli Cells / cytology,  metabolism
Spermatogonia / cytology*
Stem Cells / cytology*
Testis / cytology,  metabolism

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