Volume 5, Issue 4 (Nov 2017)
Res Mol Med (RMM) 2017, 5(4): 31-40 |
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Samareh Salavati Pour M, Hoseinpoor Kasgari F, Farsinejad A, Fatemi A, Mirzaee Khalilabadi R. Platelet-derived Microparticles Increase the Expression of hTERT Gene in Umbilical Cord Mesenchymal Stem Cells. Res Mol Med (RMM) 2017; 5 (4) :31-40
URL: http://rmm.mazums.ac.ir/article-1-262-en.html
URL: http://rmm.mazums.ac.ir/article-1-262-en.html
Maryam Samareh Salavati Pour1 
, Fatemeh Hoseinpoor Kasgari1 , Alireza Farsinejad1 , Ahmad Fatemi1 , Roohollah Mirzaee Khalilabadi 2
, Fatemeh Hoseinpoor Kasgari1 , Alireza Farsinejad1 , Ahmad Fatemi1 , Roohollah Mirzaee Khalilabadi 2
1- Department of Hematology and Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciences, Kerman, Iran
2- Department of Hematology and Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciences, Kerman, Iran , khalilabadi60@gmail.com
2- Department of Hematology and Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciences, Kerman, Iran , khalilabadi60@gmail.com
Abstract: (2976 Views)
Background: Mesenchymal stem cells have been widely considered in clinical researches because of their self-renewality and differentiation into various tissues. Nevertheless, their limited in vitro life span, which occurs only after several divisions, makes some changes in these cells, which affects all of their characteristics and remarkably reduces their application. In this study, the effect of platelet-derived microparticles, a rich source of growth factors, proteins, enzymes and microRNAs, was evaluated on the expression of hTERT gene as one of the main factors, involved in aging process and cell longevity.
Materials and methods: Umbilical cord-mesenchymal stem cells were used for this study. The cells were confirmed by evaluating their morphology and surface markers using inverted microscope and flow cytometry, respectively. Platelet microparticles were prepared by centrifuging platelet bags with different speeds, and their concentration was determined by Bradford assay. When confluency of cultured MSCs was 30%, cells were treated with 50 µg/mL of microparticles for 5 days, Then, RNA was extracted and cDNA was synthesized. The quantitative expression of hTERT gene was assessed using Real-Time PCR.
Results: The fibroblast-like cells were isolated from umbilical cord tissue, and their mesenchymal markers were approved by flowcytometry. The results of Real-Time PCR showed that the expression of hTERT gene was increased more than 3 times compared with control group.
Conclusion: It can be concluded that platelet-derived microparticles can potentially be recognized as a suitable, safe and effective method in increasing the hTERT gene expression and maybe life span of mesenchymal stem cells; however, further investigations is needed.
Materials and methods: Umbilical cord-mesenchymal stem cells were used for this study. The cells were confirmed by evaluating their morphology and surface markers using inverted microscope and flow cytometry, respectively. Platelet microparticles were prepared by centrifuging platelet bags with different speeds, and their concentration was determined by Bradford assay. When confluency of cultured MSCs was 30%, cells were treated with 50 µg/mL of microparticles for 5 days, Then, RNA was extracted and cDNA was synthesized. The quantitative expression of hTERT gene was assessed using Real-Time PCR.
Results: The fibroblast-like cells were isolated from umbilical cord tissue, and their mesenchymal markers were approved by flowcytometry. The results of Real-Time PCR showed that the expression of hTERT gene was increased more than 3 times compared with control group.
Conclusion: It can be concluded that platelet-derived microparticles can potentially be recognized as a suitable, safe and effective method in increasing the hTERT gene expression and maybe life span of mesenchymal stem cells; however, further investigations is needed.
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