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Comparison of the efficacy of three concentrations of retinoic acid for transdifferentiation induction in sheep marrow‐derived mesenchymal stem cells into male germ cells
Author(s) -
GhasemzadehHasankolaei M.,
Eslaminejad M.B.,
Batavani R.,
SedighiGilani M.
Publication year - 2014
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.12037
Subject(s) - transdifferentiation , retinoic acid , mesenchymal stem cell , immunocytochemistry , andrology , biology , alkaline phosphatase , stem cell , bone marrow , in vitro , immunology , cell culture , endocrinology , microbiology and biotechnology , medicine , biochemistry , enzyme , genetics
Summary Recent studies have shown the unique role of retinoic acid ( RA ) in the induction of transdifferentiation in mesenchymal stem cells ( MSC s) into germ cells ( GC s). This study is the first study that compares the efficacy of three different concentrations of RA for the production of male GC s in vitro . Male sheep marrow‐derived MSC s ( MMSC s) were treated with the following concentrations of RA : 1 μ m ( RA 1), 5 μ m ( RA 2) and 10 μ m ( RA 3) for a period of 21 days. The production of male GC s was evaluated by the assessment of expressions of GC ‐specific markers (by RT ‐ PCR , q RT ‐ PCR and immunocytochemistry), morphological characteristics and changes in alkaline phosphatase ( ALP ) activity. All three concentrations created male GC features. RA treatment upregulated the expressions of VASA and beta1 INTEGRIN and downregulated PIWIL2 and OCT4 . DAZL was not expressed by RA treatment. Interestingly, immunocytochemistry detected PGP 9.5 expression in all treatment groups, with the highest expression noted in the RA 3 group ( P < 0.05). GC ‐like cells along with increased ALP activity were observed in all treated cultures, too. Finally, results showed that 10 μ m RA has the most efficiency for transdifferentiation induction in MMSC s and production of male GC s in vitro .