Comparison of the efficacy of bone morphogenetic protein-4 on in vitro differentiation of murine adipose and bone marrow mesenchymal stem cells into primordial germ cells
Maryam Hosseinzadeh Shirzeyli1, Ali Tayyebiazar2, Fereshteh Aliakbari3, Fahimeh Ghasemi4, Fatemeh Eini5, Farhad Hosseinzadeh Shirzeyli6, Elmira Vanaki7, Aligholi Sobhani8
1 Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, I.R. Iran
2 Clinical Research Development Unit of Imam Khomeini Hospital, Urmia University of Medical Sciences, Urmia, I.R. Iran
3 Infertility and Reproductive Research Center, Shahid Beheshti University of Medical Sciences, Tehran, I.R. Iran
4 Cellular and Molecular Research Center; Department of Medical Biotechnology, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, I.R. Iran
5 Fertility and Infertility Research Center, Hormozgan University of Medical Science, Bandar Abbas, I.R. Iran
6 Department of Animal Science, Faculty of Agriculture and Natural Resources, Science and Research Branch, Islamic Azad University, Tehran, I.R. Iran
7 Department of Animal and Marine Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, I.R. Iran
8 Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Science, Tehran, I.R. Iran
Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Science, Tehran
Maryam Hosseinzadeh Shirzeyli
Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran
Source of Support: None, Conflict of Interest: None
Background and purpose: In vitro development of functional gametes from pluripotent stem cells is a promising prospect to treat infertility. Mesenchymal stem cells with a high degree of plasticity and less tumorigenicity are a reliable source of stem cells for the generation of gametes. The present study aimed to compare the differentiation potential in the mesenchymal stem cells that are derived from bone marrow (BMD- MSCs) and adipose tissue-derived mesenchymal stem cells (AD-MSCs) into germ cells in a culture medium containing bone morphogenic protein-4 (BMP-4).
Experimental approach: In this study, MSCs were isolated from both bone marrow and adipose tissue of murine samples. To further verify the nature of the harvested stem cells, their multipotency and surface marker were examined. The identified stem cells were cultured in a medium supplemented with 0 and 25 ng/mL of BMP-4 for 4 days. Flow cytometry analysis, immunofluorescence staining, and real RT-PCR were used to assess the expression levels in germ cell-specific biomarkers (Mvh, Dazl, Stra8, and Scp3).
Findings/Results: CD44+, CD45-, CD31-, BMD-MSCs, and AD-MSCs showed to be capable of differentiating to osteo-adipogenic lineages. The flow cytometry, immunofluorescence, and RT-PCR results indicated that early germ cell markers (Mvh and Dazl) were expressed in both types of cells but they were significantly higher in BMD-MSCs than AD-MSCs.
Conclusion and implications: Based on our results, the addition of exogenous BMP4 to the culture medium could differentiate BMD-MSCs and AD-MSCs into primordial germ cells, but it is inadequate to further develop into late germ cells in vitro. Moreover, the results revealed that, although AD-MSCs were easier to collect and had faster growth and proliferation rates than BMD-MSCs, the BMD-MSCs were better capable of differentiation into primordial germ cells. They may serve to be considered a more suitable source of MSC for in vitro generation of gametes than AD-MSCs.