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Study the effect of RG108 small molecule on neural differentiation of adipose tissue-derived stem cells
Bita Mirzaei,1,*Masoumeh Fakhr Taha ,2Arash Javeri ,3Arezoo Bazargani,4
1. Department of Stem Cells and Regenerative Medicine, Institute for Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran 2. Department of Stem Cells and Regenerative Medicine, Institute for Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran 3. Department of Stem Cells and Regenerative Medicine, Institute for Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran 4. Department of Stem Cells and Regenerative Medicine, Institute for Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran
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Introduction: Recent findings suggest the possibility of using epigenetic modifiers to reprogram somatic cells and to increase differentiation potential of stem cells. In the present study, small molecule RG108 was used to improve neural differentiation of adipose tissue-derived stem cells (ADSCs).
Methods: Human ADSCs were isolated enzymatically from adipose tissue samples obtained by abdominoplasty and were characterized. The cells were treated with concentrations of 0, 5, 25, and 50 µM RG108 for three days. Then, the effect of different concentrations of RG108 on proliferation rate and cell cycle progression was evaluated by MTT assay and flow cytometry. Also, the expression of OCT4 pluripotency marker was evaluated by flow cytometry. Finally, after 3 days of RG108 treatment, the cells were differentiated for 10 days in a medium consisting of DMEM, 1% FBS, B27, BHA, KCL, forskolin, valproic acid, hydrocortisone, and insulin. Ten days after the initiation of differentiation, the expression of PAX6, NESTIN, NSE, NeuN, NEFL mRNAs and TUJ1, MAP2, TH, proteins was evaluated.
Results: According to flow cytometry results, CD73, CD90, and CD105 markers were expressed in more than 99% of the ADSCs and the cells showed adipogenic and osteogenic differentiation. Treatment with 5 µM RG108 decreased proliferation rate and frequency of the cells in S phase of cell cycle and upregulated the expression of OCT4 pluripotency gene. Also, treatment with 5 µM RG108 before neural differentiation increased the expression of PAX6, NESTIN, NSE, NeuN, NEFL mRNAs and TUJ1, MAP2 and TH proteins.
Conclusion: The findings of the present study indicated the importance of RG108 small molecule in improvement of neural differentiation of ADSCs which may be beneficial for application of these cells in cell therapy of neurodegenerative diseases.
