Factors supporting long-term culture of bovine male germ cells.

• Sahare, Mahesh

• Kim, Sung-Min

• Otomo, Ayagi

• Komatsu, Kana

• Minami, Naojiro

• Yamada, Masayasu

• 今井, 裕

抄録

Spermatogonial stem cells (SSCs) are unipotent in nature, but mouse SSCs acquire pluripotency under the appropriate culture conditions. Although culture systems are available for rodent and human germ-cell lines, no proven culture system is yet available for livestock species. Here, we examined growth factors, matrix substrates and serum-free supplements to develop a defined system for culturing primitive germ cells (gonocytes) from neonatal bovine testis. Poly-L-lysine was a suitable substrate for selective inhibition of the growth of somatic cells and made it possible to maintain a higher gonocyte : somatic cell ratio than those maintained with gelatin, collagen or Dolichos biflorus agglutinin (DBA) substrates. Among the serum-free supplements tested in our culture medium, knockout serum replacement (KSR) supported the proliferation and survival of gonocytes better than the supplements B-27 and StemPro-SFM after sequential passages of colonies. Under our optimised culture conditions consisting of 15% KSR supplement on poly-L-lysine-coated dishes, the stem-cell and germ-cell potentials of the cultured gonocytes were maintained with normal karyotype for more than 2 months (over 13 passages). The proposed culture system, which can maintain a population of proliferating bovine germ stem cells, could be useful for studying SSC biology and germline modifications in livestock animals.

収録刊行物

• Reproduction, fertility, and development

  Reproduction, fertility, and development 2015-07-16

  CSIRO publishing

キーワード

• GDNF
• gonocytes
• self-renewal
• KSR
• long-term culture
• testis

詳細情報

• CRID: 1050001335818825472
• NII論文ID: 120005678201
• NII書誌ID: AA10718189
• ISSN: 10313613
• HANDLE: 2433/202607
• 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