Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Development of In Vitro Culture System for Male Germline Stem Cells in Porcine

돼지 웅성 생식선 줄기세포의 체외배양기법 개발

  • Kim, Yong-Hee (Department of Animal Science & Technology, Chung-Ang University) ;
  • Kim, Byung-Gak (Department of Animal Science & Technology, Chung-Ang University) ;
  • Lee, Yong-An (Department of Animal Science & Technology, Chung-Ang University) ;
  • Kim, Bang-Jin (Department of Animal Science & Technology, Chung-Ang University) ;
  • Kim, Ki-Jung (Department of Animal Science & Technology, Chung-Ang University) ;
  • Lee, Myeung-Sik (Hanwoo Experiment Station, National Institute of Animal Science, RDA) ;
  • Im, Gi-Sun (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Ryu, Buom-Yong (Department of Animal Science & Technology, Chung-Ang University)
  • 김용희 (중앙대학교 동물자원과학과) ;
  • 김병각 (중앙대학교 동물자원과학과) ;
  • 이용안 (중앙대학교 동물자원과학과) ;
  • 김방진 (중앙대학교 동물자원과학과) ;
  • 김기중 (중앙대학교 동물자원과학과) ;
  • 이명식 (농촌진흥청 축산과학원 한우개량사업소) ;
  • 임기순 (농촌진흥청 축산과학원 동물바이오공학과) ;
  • 류범용 (중앙대학교 동물자원과학과)
  • Published : 2009.09.30
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Abstract

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 $10{\sim}14$-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 $O_2$ 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, $37^{\circ}C$ temperature, and $O_2$ 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.

Keywords

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