Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Identification of a Technique Optimized for the Isolation of Spermatogonial Stem Cells from Mouse Testes

  • Han, Na Rae (Department of Animal Life Science, Kangwon National University) ;
  • Park, Hye Jin (Department of Animal Life Science, Kangwon National University) ;
  • Lee, Hyun (Department of Animal Life Science, Kangwon National University) ;
  • Yun, Jung Im (Institute of Animal Resources, Kangwon National University) ;
  • Choi, Kimyung (Optipharm Incorporation) ;
  • Lee, Eunsong (College of Veterinary Medicine, Kangwon National University) ;
  • Lee, Seung Tae (Department of Animal Life Science, Kangwon National University)
  • Received : 2018.10.24
  • Accepted : 2018.12.18
  • Published : 2018.12.31
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Abstract

To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.

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References

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