Unstable Side Population Phenotype of Mouse Spermatogonial Stem Cells In Vitro

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  • SHINOHARA Takashi
    Department of Molecular Genetics, Graduate School of Medicine, Kyoto University Japan Science and Technology Agency, CREST
  • ISHII Kei
    Department of Molecular Genetics, Graduate School of Medicine, Kyoto University
  • KANATSU-SHINOHARA Mito
    Department of Molecular Genetics, Graduate School of Medicine, Kyoto University

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Abstract

Stem cells of the side population (SP) phenotype are found in many self-renewing tissues and can be identified by their unique ability to effectively exclude the dye Hoechst 33342. We previously established a method for expanding spermatogonial stem cells (SSCs) in vitro, but the frequency of SSCs is only about 1 to 2%, limiting detailed SSC analyses. In this study, we sought to isolate SSCs from in vitro cultures by exploiting their ability to exclude Hoechst 33342. In contrast to the findings of previous in vivo studies, we found that SP cells developed in a stochastic manner in vitro. Moreover, SP cells in culture were not enriched in SSCs, but they were interconvertible with non-SP cells. Although SP cells were consistently found in testes after transplantation of cultured cells, they were not enriched in SSCs. These results show that SSCs have an unstable SP phenotype and provide evidence that SSCs change their phenotype characteristics in response to their microenvironment. <br>

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