돼지 웅성 생식선 줄기세포의 체외배양기법 개발


Development of In Vitro Culture System for Male Germline Stem Cells in Porcine

김용희, 김병각, 이용안, 김방진, 김기중, 이명식, 임기순, 류범용

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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 10~14-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 O2 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℃ temperature, and O2 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.


 재료 및 방법
  돼지 정소 세포회수
  돼지 정원줄기세포 순수도 증진
  체외 배양
  공배양세포의 준비
  공배양세포의 처리 효과
  세포배양액의 처리 효과
  배양 온도 및 혈청 첨가 효과
  배양 시 산소 농도 효과
  돼지 생식선 줄기세포의 장기간 배양


  • 김용희 Yong-Hee Kim. 중앙대학교 동물자원과학과
  • 김병각 Byung-Gak Kim. 중앙대학교 동물자원과학과
  • 이용안 Yong-An Lee. 중앙대학교 동물자원과학과
  • 김방진 Bang-Jin Kim. 중앙대학교 동물자원과학과
  • 김기중 Ki-Jung Kim. 중앙대학교 동물자원과학과
  • 이명식 Myeung Sik Lee. 농촌진흥청 축산과학원 한우개량사업소
  • 임기순 Gi-Sun Im. 농촌진흥청 축산과학원 동물바이오공학과
  • 류범용 Buom-Yong Ryu. 중앙대학교 동물자원과학과


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