Mir-106b Cluster Regulates Primordial Germ Cells Differentiation from Human Mesenchymal Stem Cells

(Pages: 294-302)
Sadaf Mahboudi, Ph.D., 1Kazem Parivar, Ph.D., 1,*Zohreh Mazaheri, Ph.D., 2Shiva Irani, Ph.D., 1
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Basic Medical Sciences Research Center, Histogenotech Company, Tehran, Iran
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Basic Medical Sciences Research Center, Histogenotech Company, Tehran, Iran
*Corresponding Address: P.O.Box: 14515/775 Department of Biology Science and Research Branch Islamic Azad University Tehran Iran Email:kazem_parivar@yahoo.com
The Cell Journal (Yakhteh) is an open access journal which means the articles are freely available online for any individual author to download and use the providing address. The journal is licensed under a Creative Commons Attribution-Non Commercial 3.0 Unported License which allows the author(s) to hold the copyright without restrictions that is permitting unrestricted use, distribution, and reproduction in any medium provided the original work is properly cited. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Mahboudi Sadaf, Parivar Kazem, Mazaheri Zohreh, Irani Shiva. Mir-106b Cluster Regulates Primordial Germ Cells Differentiation from Human Mesenchymal Stem Cells. Cell J. 2021; 23(3): 294-302.

Abstract

Objective

Numerous evidence indicates that microRNAs (miRNAs) are critical regulators in the spermatogenesis process. The aim of this study was to investigate Mir-106b cluster regulates primordial germ cells (PGCs) differentiation from human mesenchymal stem cells (MSCs).

Materials and Methods

In this experimental study, samples containing male adipose (n: 9 samples- age: 25-40 years) were obtained from cosmetic surgeries performed for the liposuction in Imam Khomeini Hospital. The differentiation of MSCs into PGCs was accomplished by transfection of a lentivector expressing miR-106b. The transfection of miR- 106b was also confirmed by the detection of a clear green fluorescent protein (GFP) signal in MSCs. MSCs were treated with bone morphogenic factor 4 (BMP4) protein, as a putative inducer of PGCs differentiation, to induce the differentiation of MSCs into PGCs (positive control). After 4 days of transfection, the expression of miR-106b, STELLA, and FRAGILIS genes was evaluated by real-time polymerase chain reaction (PCR). Also, the levels of thymocyte differentiation antigen 1 (Thy1) protein was assessed by the western blot analysis. The cell surface expression of CD90 was also determined by immunocytochemistry method. The cytotoxicity of miR-106b was examined in MSCs after 24, 48, and 72 hours using the MTT assay.

Results

MSCs treated with BMP4 or transfected by miR-106b were successfully differentiated into PGCs. The results of this study also showed that the expression of miR-106b was significantly increased after 48 hours from transfection. Also, we showed STELLA, FARGILIS, as well as the protein expression of Thy1, was significantly higher in MSCs transfected by lentivector expressing miR-106b in comparison with MSCs treated with BMP4 (P≤0.05). MTT assay showed miR-106b was no toxic during 72 hours in 1 µg/ml dose, that this amount could elevated germ cells marker significantly higher than other experimental groups (P≤0.05).

Conclusion

According to this findings, it appears that miR-106b plays an essential role in the differentiation of MSCs into PGCs.