Enrichment of Spermatogonial Stem Cells using Side Population in Teleost

• Makoto Hayashi

• Masanao Sato

• Yasuhiko Nagasaka

• Sakiko Sadaie

• Satoru Kobayashi

• Yoshizaki, Goro

抄録

Spermatogenesis originates from a small population of spermatogonial stem cells; this population can maintain continuous sperm production throughout the life of fish via self-renewal and differentiation. Despite their biological importance, spermatogonial stem cells are not thoroughly characterized because they are difficult to distinguish from their progeny 5 cells that become committed to differentiation. We previously established a novel technique for germ cell transplantation to identify spermatogonial stem cells based on their colonizing activity and their ability to initiate donor-derived gametogenesis in the rainbow trout (Oncorhynchus mykiss). Although spermatogonial stem cells can be retrospectively identified after transplantation, there is currently no technique to prospectively enrich for or purify spermatogonial stem cells. Here, we describe a method for spermatogonial stem-cell enrichment using side-population. With optimized Hoechst 33342 staining conditions, we successfully identified side-population cells among type A spermatogonia. Side-population cells were transcriptomically and morphologically distinct from non-side-population cells. To functionally determine whether the transplantable spermatogonial stem cells were enriched in the side-population fraction, we compared the colonization activity of side-population cells with that of non-side-population cells. Colonization efficiency was significantly higher with side-population cells than with non-side-population cells or with total type A spermatogonia. In addition, side-population cells could produce billions of sperm in recipient. These results indicated that transplantable spermatogonial stem cells were enriched in the side-population fraction. This method will provide biological information that may advance our understanding of spermatogonial stem 20 cells in teleosts. Additionally, this technique will increase the efficiency of germ-cell transplantation used in surrogate broodstock technology.

source:https://doi.org/10.1095/biolreprod.113.114140

source:https://kaken.nii.ac.jp/ja/grant/KAKENHI-PLANNED-25114005/

source:https://nrid.nii.ac.jp/ja/nrid/1000070281003/

identifier:25114005

収録刊行物

• Biology of Reproduction

  Biology of Reproduction 91 (1), 2014-07-01

  Oxford University Press

キーワード

• fish
• flow cytometry
• side population
• Hoechst 33342 dye
• spermatogonial stem cells
• germ cell transplantation
• 科学研究費研究成果
• サケ科魚類の進化に伴うGSC制御機構の変化
• Evolution of GSC regulation in salmonids Planned

詳細情報

• CRID: 1050001202567227904
• NII論文ID: 120006547607
• NII書誌ID: AA00565842
• ISSN: 00063363
• Web Site: http://id.nii.ac.jp/1342/00001609/
• 本文言語コード: en
• 資料種別: journal article
• データソース種別:
  • IRDB
  • CiNii Articles