Korean J Reprod Med.  2008 Mar;35(1):39-48.

Establishment and Characterization of Multipotent Germ Line Stem Cells (MGSCs) from Neonatal Mouse Testis

Affiliations
  • 1Laboratory of Reproductive Biology and Infertility, Cheil General Hospital & Women's Healthcare Center, Kwandong University College of Medicine, Seoul, Korea. junjh55@hanmail.net
  • 2Department of Medicine, Cheil General Hospital & Women's Healthcare Center, Kwandong University College of Medicine, Seoul, Korea.

Abstract


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.

Keyword

Multipotent germ line stem cells (MGSCs); Neonatal mouse testis; Embryonic stem cells (ESCs); Differentiation; Regenerative medicine

MeSH Terms

Alkaline Phosphatase
Animals
Biotechnology
Electrons
Embryoid Bodies
Embryonic Stem Cells
Feeder Cells
Germ Cells
Germ Layers
Immunohistochemistry
Karyotype
Karyotyping
Mice
Mice, Inbred ICR
Mitomycin
Multipotent Stem Cells
Regenerative Medicine
Stem Cells
Testis
Alkaline Phosphatase
Mitomycin
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