Past Issue

Volume 20, Number 4, Jan-Mar(Winter) 2019, Serial Number: 80 Pages: 505-512

Indirect Co-Culture of Testicular Cells with Bone Marrow Mesenchymal Stem Cells Leads to Male Germ Cell-Specific Gene Expressions


Mehrdad Ghorbanlou, M.Sc, 1, Alireza Abdanipour, Ph.D, 1, *, Reza Shirazi, Ph.D, 2, 3, Nasim Malekmohammadi, M.Sc, 1, Saeed Shokri, Ph.D., 1, Reza Nejatbakhsh, Ph.D, 1, *,
Department of Anatomical Sciences, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran
*Corresponding Address: P.O.Box: 4513956111 Department of Anatomical Sciences School of Medicine Zanjan University of Medical Sciences Zanjan Iran Emails:abdani.anatomy@yahoo.com,reza_nejat@yahoo.com

Abstract

Objective

Non-obstructive azoospermia is mostly irreversible. Efforts to cure this type of infertility have led to the application of stem cells in the reproduction field. In the present study, testicular cell-mediated differentiation of male germ-like cells from bone marrow-derived mesenchymal stem cells (BM-MSCs) in an in vitro indirect co-culture system is investigated.

Materials and Methods

In this experimental study, mouse BM-MSCs were isolated and cultured up to passage three. Identification of the cells was evaluated using specific surface markers by flow-cytometry technique. Four experimental groups were investigated: control, treatment with retinoic acid (RA), indirect co-culture with testicular cells, and combination of RA and indirect co-culture with testicular cells. Finally, following differentiation, the quantitative expression of germ cell-specific markers including Dazl, Piwil2 and Stra8 were evaluated by real-time polymerase chain reaction (PCR).

Results

Molecular analysis revealed a significant increase in Dazl expression in the indirect co-culture with testicular cells group in comparison to the control group. Quantitative expression level of Piwil2 was not significantly changed in comparison to the control group. Stra8 expression was significantly higher in RA group in comparison to other groups.

Conclusion

Indirect co-culture of BM-MSCs in the presence of testicular cells leads to expression of male germ cell-specific gene, Dazl, in the induced cells. Combination of co-culture with testicular cells and RA did not show any positive effect on the specific gene expressions.