Differentiation of bone marrow derived mesenchymal stem cells into male germ-like cells in co-culture with testicular cells

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Objective. Stem cell therapy, specifically, pre-induction of mesenchymal stem cells toward male germ-like cells may be useful in patients with azoospermia. The aim of this study was to evaluate in vitro differentiation of mouse bone marrow-derived mesenchymal stem cells (BMSCs) into male germ-like cells by indirect co-culture with testicular cells in the presence of bone morphogenetic protein 4 (BMP4).

Methods. Experimental groups included: control (mouse BMSCs), treatment group-1 (BMSCs treated with BMP4), treatment group-2 (indirect co-culture of BMSCs with mouse testicular cells in the presence of BMP4) and treatment group-3 (indirect co-culture of BMSCs with testicular cells). BMSCs-derived male germ-like cells were evaluated by the expression of Dazl, and Stra8 using RT-qPCR.

Results. Stra8 gene expression was significantly increased in the treatment group-2 and Dazl gene was significantly increased in the treatment group-1 compared to other groups. In conclusion, indirect co-culturing of BMSCs with testicular cells and BMP4 leads to the differentiation of BMSCs into male germ-like cells which express specific male germ-like genes. Testicular cells released factors that contributed to the differentiation of BMSCs into male germ progenitor cells.

Conclusion. This study suggests that mesenchymal stem cells may be differentiated into male germ-like cells and therefore, may be a novel treatment option for men with azoospermia.

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  • de Rooij DG. Proliferation and differentiation of spermatogonial stem cells. Reproduction 121 347–354 2001.

  • de Sousa Lopes SM Roelen BA Monteiro RM Emmens R Lin HY Li E Lawson KA Mummery CL. BMP signaling mediated by ALK2 in the visceral endoderm is necessary for the generation of primordial germ cells in the mouse embryo. Genes Dev 18 1838–1849 2004.

  • Dudley BM Runyan C Takeuchi Y Schaible K Molyneaux K. BMP signaling regulates PGC numbers and motility in organ culture. Mech Dev 124 68–77 2007.

  • Fabbri A Knox G Buczko E Dufau ML. β-endorphin production by the fetal Leydig cell: Regulation and implications for paracrine control of Sertoli cell function. Endocrinology 122 749–755 1998.

  • Foresta C Moro E Ferlin A. Y chromosome microdeletions and alterations of spermatogenesis. Endocr Rev 22 226–239 2001.

  • Geens M Sermon KD Van de Velde H Tournaye H. Sertoli cell-conditioned medium induces germ cell differentiation in human embryonic stem cells. J Assist Reprod Genet 28 471–480 2011.

  • Gimble JM Katz AJ Bunnell BA. Adipose-derived stem cells for regenerative medicine. Circ Res 100 1249–1260 2007.

  • Hamidabadi HG Pasbakhsh P Amidi F Soleimani M Forouzandeh M Sobhani A. Functional concentrations of BMP4 on differentiation of mouse embryonic stem cells to primordial germ cells. Int J Fertil Steril 5 104–109 2011.

  • Hansson V Weddington SC McLean WS Smith AA Nayfeh SN French FS Ritzen EM. Regulation of seminiferous tubular function by FSH and androgen. J Reprod Fertil 44 363–375 1975.

  • Hussein A. Evaluation of diagnostic testis biopsy and the repetition of testicular sperm extraction surgeries in infertility patients. Fertil Steril 100 88–93 2013.

  • Iammarrone E Balet R Lower AM Gillott C Grudzinskas JG. Male infertility. Best Pract Res Clin Obstet Gynaecol 17 211–229 2003.

  • Lacham-Kaplan O Chy H Trounson A. Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes. Stem Cells 24 266–273 2006.

  • Latifpour M Shakiba Y Amidi F Mazaheri Z Sobhani A. Differentiation of human umbilical cord matrix-derived mesenchymal stem cells into germ-like cells. Avicenna J Med Biotechnol 6 218–227 2014.

  • Lee JH Engel W Nayernia K. Stem cell protein Piwil2 modulates expression of murine spermatogonial stem cell expressed genes. Mol Reprod Dev 73 173–179 2006.

  • Makoolati Z Movahedin M Forouzandeh-Moghadam M. In vitro germ cell differentiation from embryonic stem cells of mice: induction control by BMP4 signalling. Biosci Rep 36 pii: e00407 2016.

  • Margioris AN Liotta AS Vaudry H Bardin CW Krieger DT. Characterization of immunoreactive proopiomelanocortin-related peptides in rat testes. Endocrinology 113 663–671 1983.

  • Miryounesi M Nayernia K Dianatpour M Mansouri F Modarressi MH. Co-culture of mouse embryonic stem cells with Sertoli cells promote in vitro generation of germ cells. Iran J Basic Med Sci 16 779–783 2013.

  • Pellegrini M Grimaldi P Rossi P Geremia R Dolci S. Developmental expression of BMP4/ALK3/SMAD5 signaling pathway in the mouse testis: a potential role of BMP4 in spermatogonia differentiation. J Cell Sci 116 3363–3372 2003.

  • Saitou M Barton SC Surani MA. A molecular programme for the specification of germ cell fate in mice. Nature 418 293–300 2002.

  • Saitou M Payer B Lange UC Erhardt S Barton SC Surani MA. Specification of germ cell fate in mice. Philos Trans R Soc Lond B Biol Sci 358 1363–1370 2003.

  • Shirazi R Zarnani AH Soleimani M Abdolvahabi MA Nayernia K Ragerdi Kashani I. BMP4 can generate primordial germ cells from bone-marrow-derived pluripotent stem cells. Cell Biol Int 36 1185–1193 2012.

  • Silva C Wood JR Salvador L Zhang Z Kostetskii I Williams CJ Strauss JF 3rd. Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone. Mol Reprod Dev 76 11–21 2009.

  • Singh A Jahan N Radhakrishnan G Banerjee BD. To evaluate the efficacy of combination antioxidant therapy on oxidative stress parameters in seminal plasma in the male infertility. J Clin Diagn Res 10 QC14–QC17 2016.

  • Skinner MK Fritz IB. Testicular peritubular cells secrete a protein under androgen control that modulates Sertoli cell functions. Proc Natl Acad Sci U S A 82 114–118 1985.

  • Skinner MK. Cell-cell interactions in the testis. Endocr Rev 12 45–77 1991.

  • Surani MA Ancelin K Hajkova P Lange UC Payer B Western P Saitou M. Mechanism of mouse germ cell specification: a genetic program regulating epigenetic reprogramming. Cold Spring Harb Symp Quant Biol 69 1–9 2004.

  • Vannelli BG Barni T Orlando C Natali A Serio M Balboni GC. Insulin-like growth factor-1 (IGF-I) and IGF-I receptor in human testis: an immunohistochemical study. Fertil Steril 49 666–669 1988.

  • Walker WH Cheng J. FSH and testosterone signaling in Sertoli cells. Reproduction 130 15–28 2005.

  • West FD Roche-Rios MI Abraham S Rao RR Natrajan MS Bacanamwo M Stice SL. KIT ligand and bone morphogenetic protein signaling enhances human embryonic stem cell to germ-like cell differentiation. Hum Reprod 25 168–178 2010.

  • Xie L Lin L Tang Q Li W Huang T Huo X Liu X Jiang J He G Ma L. Sertoli cell-mediated differentiation of male germ cell-like cells from human umbilical cord Wharton’s jelly-derived mesenchymal stem cells in an in vitro co-culture system. Eur J Med Res 20 9 2015.

  • Yang Y Feng Y Feng X Liao S Wang X Gan H Wang L Lin X Han C. BMP4 cooperates with retinoic acid to induce the expression of differentiation markers in cultured mouse spermatogonia. Stem Cells Int 2016 9536192 2016.

  • Yu Z Ji P Cao J Zhu S Li Y Zheng L Chen X Feng L. Dazl promotes germ cell differentiation from embryonic stem cells. J Mol Cell Biol 1 93–103 2009.

  • Zhang D Liu X Peng J He D Lin T Zhu J Li X Zhang Y Wei G. Potential spermatogenesis recovery with bone marrow mesenchymal stem cells in an azoospermic rat model. Int J Mol Sci 15 13151–13165 2014.

  • Zhou Q Nie R Li Y Friel P Mitchell D Hess RA Small C Griswold MD. Expression of stimulated by retinoic acid gene 8 (Stra8) in spermatogenic cells induced by retinoic acid: an in vivo study in vitamin A-sufficient postnatal murine testes. Biol Reprod 79 35–42 2008.

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