Cluster Regulates Primordial Germ Cells Differentiation from Human Mesenchymal Stem Cells.

[OBJECTIVE] Numerous evidence indicates that microRNAs (miRNAs) are critical regulators in the spermatogenesis process. The aim of this study was to investigate 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 . The transfection of 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 , , and 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 was examined in MSCs after 24, 48, and 72 hours using the MTT assay.

[RESULTS] MSCs treated with BMP4 or transfected by 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 as well as the protein expression of Thy1, was significantly higher in MSCs transfected by lentivector expressing in comparison with MSCs treated with BMP4 (P≤0.05). MTT assay showed 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 plays an essential role in the differentiation of MSCs into PGCs.