초록

Ⅳ. 연구개발결과
1) 생쥐를 이용한 정소조직의 체외배양 기술개발
2) 돼지 정소조직의 체외 배양 조건 확립
3) 정자형성과정 유도 효율 증진을 위한 첨가제 효과 분석
4) 돼지 정소로부터 정원줄기세포의 회수와 순수도 증진 기법의 최적화
5) 돼지 정소조직의 이용성 증진을 위한 효율적인 동결보존 기술 개발
6) 돼지 정원줄기세포의 유전자 도입법 최적화
7) 체외 배양된 정소조직으로부터 정자 생산 및 형질전환 수정란 생산유도

Abstract ▼

Spermatogonial stem cells (SSCs) is the germ-line stem cell that is the foundation of spermatogenesis within testis. These have unique abilities to self-renew and differentiate into committed cells that can finally produce sperm. Especially, SSCs in males are the vehicle through which genetic inform

Spermatogonial stem cells (SSCs) is the germ-line stem cell that is the foundation of spermatogenesis within testis. These have unique abilities to self-renew and differentiate into committed cells that can finally produce sperm. Especially, SSCs in males are the vehicle through which genetic information is transmitted to the next generation for mammals. Therefore SSCs have been the focus of one of important sources for germline-modification to generate transgenic. Thus genetic modification at the level of SSC is thought to be a relevant method that is possible to transmit genetic information from parents to the progeny.
The object of this study is to improve the technology for producing transgenic animal using porcine male germ-line stem cells and in vitro testis tissue culture system and ultimately to develop the practical application for the production of disease-model animal, bioreactor animal.
The main results of this research were as follows:
1) Evaluated in vitro testis tissue culture system in mice
2) Established in vitro testis tissue culture system in pigs
3) Investigated the effects of different environmental factors
4) Developed the methods for the purification of SSC in the testis cell population
5) Investigated an efficient cryopreservation method for porcine SSCs
6) Established a condition for transducing eGFP-lentiviral vector into porcine SSCs
7) Produced sperm from in vitro cultured testis tissues transduced with eGFP and induce the production of fertilized egg