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Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes
Author(s) -
Han NR,
Park YH,
Yun JI,
Park HJ,
Park MH,
Kim MS,
Choi JH,
Lee E,
Gong SP,
Lim JM,
Lee ST
Publication year - 2014
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12351
Subject(s) - biology , homeobox protein nanog , stem cell , sox2 , microbiology and biotechnology , alkaline phosphatase , andrology , stromal cell , embryonic stem cell , immunology , gene , induced pluripotent stem cell , genetics , cancer research , biochemistry , enzyme , medicine
Contents Recently, isolation and in vitro culture of putative spermatogonial stem cells ( SSC s) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSC s ( FSSC s) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSC s among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells ( TSC s) derived from cats, dogs and mice, and mouse embryonic fibroblasts ( MEF s). The formation and growth of colonies derived from SSC s cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase ( AP ) activity and expression of SSC ‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSC s from cat showed the greatest colony formation, growth and maintenance of FSSC s, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC ‐specific genes ( NANOG , OCT 4 , SOX 2 and CD 9 ). Accordingly, these results demonstrate that feline TSC s could be used as feeder cells to support the establishment and maintenance of SSC s from domestic cats.