Studying the effect of serial passages in human mesenchymal stem cells on gene expression of TERT and miR-138 genes in vitro
Studying the effect of serial passages in human mesenchymal stem cells on gene expression of TERT and miR-138 genes in vitro
Sami Beit Sayah Ozari,1Seyedeh Fatemeh Azghadi,2Amir Atashi,3,*
1. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2. Student Research Committee, School of Allied Medical Sciences, Shahroud University of Medical Sciences, Shahroud, Iran 3. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Introduction: Mesenchymal stem cells (MSCs) are of great significance in tissue engineering and disease treatment. There is a small number of these cells in the body. Therefore, their expansion is needed for clinical applications. However, in vitro expansion can shorten telomere length, cause aging in cells, and decrease their quality in cellular therapy. In this study, the goal is to assess the expression of miR-138 gene as a known telomerase regulator through consecutive MSCs passages.
Methods: MSCs isolation was conducted by RosetteSep kit from human bone marrow aspiration. After five passages, mRNA and miRNA were isolated via TRIZOL through the phenol-chloroform method. cDNA was synthesized, and the expression of miR-138 gene and TERT were evaluated through Real-Time PCR.
Results: The outcomes indicated a significant decrease in the expression of TERT gene after one passage. A notable increase in miR-138 expression was detected as well (p<0,05).
Conclusion: Evaluating the expression of miR-138 and TERT genes in expanding MSCs indicates the potential use of miR-138 in increasing the capability of in vitro MSCs expansion.