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Porcine spermatogonial stem cells self‐renew effectively in a three dimensional culture microenvironment
Author(s) -
Park Ji Eun,
Park Min Hee,
Kim Min Seong,
Park Yeo Reum,
Yun Jung Im,
Cheong Hee Tae,
Kim Minseok,
Choi Jung Hoon,
Lee Eunsong,
Lee Seung Tae
Publication year - 2017
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1002/cbin.10844
Subject(s) - microbiology and biotechnology , cell culture , biology , stem cell , in vitro , agarose , cell growth , genetics
Generally, self‐renewal of spermatogonial stem cells (SSCs) is maintained in vivo in a three‐dimensional (3D) microenvironment consisting of the seminiferous tubule basement membrane, indicating the importance of the 3D microenvironment for in vitro culture of SSCs. Here, we report a 3D culture microenvironment that effectively maintains porcine SSC self‐renewal during culture. Porcine SSCs were cultured in an agarose‐based 3D hydrogel and in 2D culture plates either with or without feeder cells. Subsequently, the effects of 3D culture on the maintenance of undifferentiated SSCs were identified by analyzing cell colony formation and morphology, AP activity, and transcriptional and translational regulation of self‐renewal‐related genes and the effects on proliferation by analyzing cell viability and single cell‐derived colony number. The 3D culture microenvironment constructed using a 0.2% (w/v) agarose‐based 3D hydrogel showed the strongest maintenance of porcine SSC self‐renewal and induced significant improvements in proliferation compared with 2D culture microenvironments. These results demonstrate that self‐renewal of porcine SSCs can be maintained more effectively in a 3D than in a 2D culture microenvironment. Moreover, this will play a significant role in developing novel culture systems for SSCs derived from diverse species in the future, which will contribute to SSC‐related research.