期刊
ANIMAL REPRODUCTION SCIENCE
卷 137, 期 1-2, 页码 45-52出版社
ELSEVIER
DOI: 10.1016/j.anireprosci.2012.12.005
关键词
Male germline stem cells (mGSC); Leukemia inhibitory factor (LIF); Extracellular matrix; Basic fibroblast growth factor (bFGF); 6-Bromoindirubin-3 '-oxime (BIO); Dairy goat
资金
- National Natural Science Foundation of China [30972097, 31272518]
- Key Program of State Education Ministry [109148]
- Program for New Century Excellent Talents in University [NCET-09-0654]
- Special Fund for Agroscientific Research in the Public Interest [201103038]
- Scientific Research Program of Shaanxi Province [2011K02-06]
- Fundamental Research Funds for the Central Universities [QN2011012]
Male germline stem cells (mGSC) reside in the basement of seminiferous tubules of the testis and have the capacity of self-renewal and differentiation into sperm throughout the life of animals. Reports on mice and human mGSC have demonstrated that mGSC are an unlimited resource of pluripotent stem cells for sperm production. The conditions of isolation and culture of mouse and human mGSC are well developed; however, the systematic culture conditions of dairy goat mGSC are still deficient although there have been several reports of successful cultures. With the present research, several key elements of isolation and culture of dairy goat mGSC have been determined. Details for the conditions of isolation of dairy testicular spermatogonium cells were optimized, and effects of several extracellular matrix types, ages of dairy goat, and cytokines on enrichment and culture of mGSC were compared. Biological characteristics of the cells were also evaluated by RT-PCR and immunofluorescent staining. The results indicated there is one kind of enzyme cocktail (CTHD (1 mg/ml collagenase, 10 mu g/ml DNase, 1 mg/ml hyaluronidase and 1 mg/ml trypsin) combined TD (0.25% trypsin and 10 mg/ml DNaseI)) that can be used to successfully isolate dairy goat testicular spermatogonium cells efficiently; and fibronectin as well as laminin were efficient extracellular matrix to enrich mGSC among the extracellular matrix types evaluated. Age of dairy goat clearly influenced the cultures of dairy goat mGSC with the efficiency of establishment of an mGSC line being greater if the age of the dairy goat is younger. Some cytokines e.g. BIO (A GSK3 inhibitor, 6-bromoindirubin-3'-oxime) and basic fibroblast growth factor (bFGF) acted positively on the maintenance of proliferation and pluripotency of mGSC. Leukemia inhibitory factor (LIF) might, however, inhibit the proliferation of dairy goat mGSC. These cultured mGSC maintained similar characteristics as mouse and human mGSC. These results provide an efficient system to isolate and culture of dairy goat mGSC. (C) 2012 Elsevier B.V. All rights reserved.
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