Daneshvar Medicine

Daneshvar Medicine

The effect of platelet gel-containing culture medium on the viability and expression of PAX6 and RPE65 genes in human retinal pigment epithelial cells

Document Type : Original Article

Authors
Maryam Amin 1
Masoud Soleimani 1
Mozhgan Rezaie Kanavi 2
Reihane Ghasemi Tarei 3
1 Department of Hematology, School of Medicine, Tarbiyat Modares University, Tehran, Iran
2 Ocular Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
3 Department of Physiology, School of Medicine, Shahed University, Tehran, Iran
10.22070/daneshmed.2020.2973
Abstract
Background and Objective: Degradation of human retinal pigment epithelium (hRPE) during some disorders such as aging is associated with severe impairment of ocular optical receptor function. Cellular and tissue transplantation of RPE layer is one of the treatment options that has been considered in recent years. The use of platelet gel has shown beneficial effects on the growth of various cultured cells.This study was conducted to investigate the effects of three different concentrations of platelet gel on the viability and the expression of PAX6 and RPE65 genes of cultivated RPE cells.
Materials and Methods: hRPE cells extracted from the human eye were cultured in plates containing platelet gel (PL) of different concentrations (10%, 20%, and 30%). Cells viability was tested by MTT test and the expression RPE65 and PAX6 genes were analyzed by using real time PCR. The number of expression changes of each gene in different treatments compared to the control and according to the internal GAPDH gene was calculated according to the 2-∆∆Ct method and the results were compared with each other in a bar chart.
Results: After 7 days, cultivated hRPE cells in culture media containing 30% PL showed a high rate of viability in comparison with those cultured in 10% PL (P<0.05). RPE65 gene expressions in cell cultures treated with 20% and 30% PL were higher than controls (P<0.05). However, there was no significant difference in the expression of PAX6 gene between the cell cultures treated with 20% and 30% platelet gels.
Conclusion: our results showed that using platelet gels in hRPE cell cultures can improve cell viability and identity, which can be promising for optimizing future clinical cell therapy strategies.
Keywords
PAX6
PRE65
Platelet gel
Retinal Pigmented Epithelium (RPE)
PCR
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Daneshvar Medicine
Volume 28, Issue 4 - Serial Number 149
September and October 2020
Pages 1-9