مقالات پذیرفته شده در هشتمین کنگره بین المللی زیست پزشکی
Evaluation of miR-183 cluster (183/96/182) expression in human adipose– derived mesenchymal stem cells (hAD-MSCs) differentiated into photoreceptor like cell under growth factor induction
Evaluation of miR-183 cluster (183/96/182) expression in human adipose– derived mesenchymal stem cells (hAD-MSCs) differentiated into photoreceptor like cell under growth factor induction
Samira Asgharzade,1,*Vahid Izadan,2
1. 1-Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran 2- Department of Molecular Medicine, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran 2. Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
Introduction: Degeneration and apoptotic demise of the photoreceptor cell layer within the retina represents a significant cause of irreversible blindness in modern medicine. Stem cell therapy stands out as a promising approach for repairing retinal damage. Moreover, external signals are crucial in directing lineage commitment, guiding fate-restricted progenitors toward photoreceptor lineage from stem cell progeny in vitro. Research indicates that taurine and retinoic acid (RA) initially influence progenitor lineage in an instructive and lineage-specific manner, promoting the development of photoreceptor-like cells from stem cells. Additionally, miRNAs are pivotal in differentiation processes, with the miR183 family notably facilitating the differentiation of neural retina cells and photoreceptors. This study seeks to explore the impact of RA and taurine on differentiating human adipose-derived mesenchymal stem cells (hAD-MSCs) into photoreceptor-like cells while assessing the expression of the miR-183 cluster (183/96/182).
Methods: hAD-MSCs were purchased from Pasteur Institute, Tehran, and cultured under standard conditions. After the three-cell passage, there are 3 groups, A) DMEM/F12 without growth factor as control) B) DMEM/F12 complemented with taurine (50 μmol/L) C) DMEM/F12 complemented with taurine (50 μmol/L) and RA (1 µM).
Subsequently, cellular change morphology was detected following 14 days under an inverted microscope. Then, the relative expression levels. Then, gene expression of photoreceptor cell biomarkers (Rho and Recoverin) and miR-183 cluster (183/96/182) were examined by quantitative polymerase chain reaction (Q-PCR).
Results: Cellular morphology demonstrated spindle elongated morphology in taurine and the combination of the taurine/retinoic acid group. The results showed that taurine and the combination of taurine/retinoic acid led to a significant increase in the expression of Rho and Recoverin genes compared to the control group (P<0.0001). the taurine group induced the expression of miR-182 (P<0.01) but there was no significant effect on mir-183 expression, and taurine and RA led to a substantial increase in the expression of miR-183 and mir182 (P<0.0001). Investigations showed that the taurine and RA had no significant effect on miR-96 expression.
Conclusion: The results of the present study show that growth factors (taurine and RA) can lead to the differentiation of hAD-MSCs into rhodopsin-positive cells and the expression of specific markers (Rho and Recoverin). Also, the increased expression of the miR-183 cluster has a key role in the differentiation pathway of photoreceptor and neuroretina cells.