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KMID : 1023620090330030171
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Reproductive and Developmental Biology
2009 Volume.33 No. 3 p.171 ~ p.177
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Development of In Vitro Culture System for Male Germline Stem Cells in Porcine
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Kim Yong-Hee
Kim Byung-Gak
Lee Yong-An
Kim Bang-Jin
Kim Ki-Jung
Lee Myeung-Sik
Im Gi-Sun
Ryu Buom-Yong
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Abstract
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Spermatogonial stem cells(SSCs) only are responsible for the generation of progeny and for the transmission of genetic information to the next generation in male. Other in vitro studies have cultured SSCs for proliferation, differentiation, and genetic modification in mouse and rat. Currently, information regarding in vitro culture of porcine Germline Stem Cell(GSC) such as gonocyte or SSC is limited and is in need of further studies. Therefore, in this study, we report development of a successful culture system for gonocytes of neonatal porcine testes. Testis cells were extracted from -day-old pigs. These cells were harvested using enzymatic digestion, and the harvested cells were purified with combination of percoll, laminin, and gelatin selection techniques. The most effective culture system of porcine gonocytes was established through trial experiments which made a comparison between different feeder cells, medium, serum concentrations, temperatures, and tensions. Taken together, the optimal condition was established using C166 or Mouse Embryonic Fibroblast(MEF) feeder cell, Rat Serum Free Medium(RSFM), 0% serum concentration, temperature, and 20% tension. Although we discovered the optimal culture condition for proliferation of porcine gonocytes, the gonocyte colonies ceased to expand after one month. These results suggest inadequate acquirement of ingredients essential for long term culture of porcine GSCs. Consequently, further study should be conducted to establish a successful long-term culture system for porcine GSCs by introducing various growth factors or nutrients.
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KEYWORD
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Spermatogonial stem cells, Germline stem cells, Gonocyte, Porcine, In vitro culture
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