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대한생식의학회지 제35권 제1호 2010년
신생 생쥐 고환에서 기인한 다분화능 생식줄기세포주의 확립 및 특성 분석
관동대학교 의과대학 제일병원 생식생물학 및 불임연구실1, 의학과2
한상철1, 송행석1,2, 전진현1,2*,
Establishment and Characterization of Multipotent Germ Line Stem Cells (MGSCs) from Neonatal Mouse Testis
Sang Chul Han1, Haengseok Song1,2, Jin Hyun Jun1,2*
1Laboratory of Reproductive Biology and Infertility and 2Department of Medicine, Cheil General Hospital & Women's Healthcare Center, Kwandong University College of Medicine, Seoul, Korea
Objective: The aim of this study was to investigate whether multipotent germline stem cells (MGSCs) can be established from neonatal mouse testis. Methods: Various cells containing MGSCs were collected from neonatal testis of ICR mice and allocated to plates for in vitro culture. After 7 days in culture, the cells were passed to a fresh culture plate and continuously cultured. From the third or fourth passage, the presumed MGSCs were cultured and maintained on mitomycin C-inactivated STO feeder cells. The MGSCs were cultured in a condition where mouse embryonic stem cells (ESCs) are cultured. Characteristics of the MGSCs were evaluated by RT-PCR, immunocytochemistry, alkaline phosphatase activity, karyotyping, and transmission electron microscopy. Results: Two MGSCs lines were established from 9 pooled sets of neonatal testicular cells. MGSCs colonies were morphologically undistinguishable from ESCs colonies and both MGSC lines as well as ESCs expressed undifferentiated stem cell markers, such as Thy-1, Oct-4, Nanog, Sox2 and alkaline phosphatase. Fine structure of undifferentiated MGSCs were similar to those of ESCs and 60% of MGSCs (12/20) had normal karyotype at passage 10. They were able to form embryoid bodies (EBs) and MGSC-derived EBs expressed marker genes of three germ layers. Conclusion: We could establish the MGSCs from neonatal mouse testis and they were differentiated to multipotent lineages of three germ layers. Molecular characteristics of MGSCs were similar to those of ESCs. Our results suggest a possibility that multipotent stem cells derived from testis, the MGSCs, could replace the ESCs in biotechnology and regenerative medicine.
키워드 : Multipotent germ line stem cells (MGSCs), Neonatal mouse testis, Embryonic stem cells (ESCs), Differentiation, Regenerative medicine
교신저자 : Ikang67@yahoo.co.kr
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