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Efficiency of adult mouse spermatogonial stem cell colony formation under several culture conditions

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Abstract

The aim of this study was to compare the in vitro effects of glial cell line-derived neurotrophic factor, stem cell factor, granulocyte macrophage-colony stimulating factor, and co-culture with Sertoli cells on the efficiency of adult mouse spermatogonial stem cells colony formation. For these purpose, both Sertoli and spermatogonial cells were isolated from adult mouse testes. The identity of the cells was confirmed through analysis of alkaline phosphatase activity, immunocytochemistry against OCT-4, c-kit, and vimentin, and also by transplantation of these cells in the recipient testes. The isolated spermatogonial cells were treated either with various concentrations of the above mentioned factors or co-cultured with Sertoli cells for 3 wk. The spermatogonial cells of the resulting colonies were transplanted via rete testis into the mouse testes, which were irradiated with 14 Gy. The results indicated that glial cell line-derived neurotrophic factor is the most appropriate factor for in vitro colonization of adult mice spermatogonial cells compared with other cytokines and growth factors. A short-term co-culture with Sertoli cells showed a significant increase in the number and diameter of the colonies compared with the treated growth factors and the control group. We have also demonstrated that mouse spermatogonial stem cells in the colonies after co-culturing with Sertoli cells could induce spermatogenesis in the recipient testes after transplantation.

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Acknowledgments

We highly appreciate the contribution of Dr. Ans van Pelt and Prof. Dr. Dirk de Rooij (Center of Reproductive Medicine, AMC, Amsterdam) for their consults and the Royan Institute (Tehran, Iran) for providing technical assistance. This work was supported by a research grant from Tarbiat Modares University (Tehran, Iran).

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  1. Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Jalale-Ale-Ahmad highway, P.O. Box: 14115-175, Tehran, Iran

    M. Koruji & M. Movahedin

  2. Department of Genetics, School of Basic Sciences, Tarbiat Modares University, Tehran, Iran

    S. J. Mowla

  3. Department of Genetics, Institute of Royan, Tehran, Iran

    H. Gourabi

  4. Department of Radiation Oncology, Shohada-E-Tajrish Hospital, Tehran, Iran

    A. J. Arfaee

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  1. M. Koruji
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  2. M. Movahedin
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Correspondence to M. Movahedin.

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Koruji, M., Movahedin, M., Mowla, S.J. et al. Efficiency of adult mouse spermatogonial stem cell colony formation under several culture conditions. In Vitro Cell.Dev.Biol.-Animal 45, 281–289 (2009). https://doi.org/10.1007/s11626-008-9169-y

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