Han N R, Park Y H, Yun J I, Park H J, Park M H, Kim M S, Choi J H, Lee E, Gong S P, Lim J M, Lee S T
Department of Animal Biotechnology, College of Animal Life and Science, Kangwon National University, Chuncheon, Korea.
Reprod Domest Anim. 2014 Oct;49(5):705-10. doi: 10.1111/rda.12351. Epub 2014 Jun 30.
Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long-term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs 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, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome-shaped morphology, AP activity and expression of SSC-specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.
最近，已经开展了家猫中假定的精原干细胞（SSCs）的分离和体外培养。然而，促进猫科动物精原干细胞（FSSCs）建立和长期维持的细胞龛条件尚未得到描述。因此，我们在FSSC龛的各种因素中，研究了用于刺激FSSCs集落形成和生长的饲养细胞类型。从猫睾丸中分离出的精原干细胞在源自猫、狗和小鼠的有丝分裂失活的睾丸基质细胞（TSCs）以及小鼠胚胎成纤维细胞（MEFs）上进行培养。在第0代时鉴定了在每种类型饲养细胞上培养的SSCs衍生集落的形成和生长情况，并在第4代时研究了存活集落的形态、碱性磷酸酶（AP）活性和SSC特异性基因的表达。在这些不同的饲养细胞中，来自猫的TSCs对FSSCs的集落形成、生长和维持表现出最强的作用，并且到第4代培养的SSC集落呈现出典型的圆顶形形态、AP活性以及SSC特异性基因（NANOG、OCT4、SOX2和CD9）的表达。因此，这些结果表明猫科动物TSCs可作为饲养细胞来支持家猫SSCs的建立和维持。
