Volume 5, Issue 4 (Nov 2017)
Res Mol Med (RMM) 2017, 5(4): 12-22 |
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Hoseinpour Kasgari F, Samareh Salavati Pour M, Farsinejad A, Fatemi A, Mirzaee Khalilabadi R. Platelet Rich in Growth Factors (PRGF): A Suitable Replacement for Fetal Bovine Serum (FBS) in Mesenchymal Stem Cell Culture. Res Mol Med (RMM) 2017; 5 (4) :12-22
URL: http://rmm.mazums.ac.ir/article-1-263-en.html
URL: http://rmm.mazums.ac.ir/article-1-263-en.html
Fatemeh Hoseinpour Kasgari1 
, Maryam Samareh Salavati Pour1 , Alireza Farsinejad1 , Ahmad Fatemi1 , Roohollah Mirzaee Khalilabadi 2
, Maryam Samareh Salavati Pour1 , Alireza Farsinejad1 , Ahmad Fatemi1 , Roohollah Mirzaee Khalilabadi 2
1- 1. Department of Hematology and Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciencs, Kerman, Iran
2- 1. Department of Hematology and Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciencs, Kerman, Iran ,khalilabadi60@gmail.com
2- 1. Department of Hematology and Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciencs, Kerman, Iran ,
Abstract: (3486 Views)
Background: Due to high differentiation potential and self-renewality, Mesenchymal Stem Cells are now widely considered by researchers in several diseases. FBS is used as a supplement in culture media for proliferation, differentiation, and other culture processes of MSCs, which is associated with transmission risk of a variety of infections as well as immune responses. PRGF derived from platelets contains growth factors causing the growth and proliferation of MSCs. This study was conducted to compare the effect of PRGF in comparison to FBS on the expression of h-TERT gene, in umbilical cord derived MSCs.
Materials and Methods: This study is an experimental research. Four expired platelet concentrate bags were obtained from Kerman blood transfusion center, and PRGF was prepared by multiple centrifugation rounds of the platelet bag. Calcium chloride was added as an anticoagulant to PRGF in order to prevent gelatinization of the culture medium. On the other hand, mesenchymal stem cells were isolated from the umbilical cord as a primary culture. The phenotype of the cells was confirmed by flow cytometry, and the cells were randomly cultured as control (using FBS) and experimental (using PRGF) groups. The expression of the gene involved in increasing cell longevity (h-TERT) was investigated by real-time PCR technique after six days.
Results: Mesenchymal stem cells were successfully isolated from the umbilical cord. Morphologically, the mesenchymal cells cultured in the experimental group (using PRGF) were similar to the cells in the control medium. The cells exhibited a high expression level of CD73, CD90, and CD105, while the surface markers of hematopoietic cells such as CD45 and CD34 were slightly expressed. Therefore, there was no significant difference in the expression of cell surface markers between control and experimental groups. In this study, using the real-time PCR technique, it was shown that PRGF derived from the platelet could increase the expression of h-TERT gene in the umbilical cord mesenchymal stem cells compared with the control.(P = 0.034)
Conclusion: PRGF have been shown to be effective in increasing expression of h-TERT gene in the umbilical cord mesenchymal stem cells and may also be an appropriate substitute for FBS in cell culture media.
Materials and Methods: This study is an experimental research. Four expired platelet concentrate bags were obtained from Kerman blood transfusion center, and PRGF was prepared by multiple centrifugation rounds of the platelet bag. Calcium chloride was added as an anticoagulant to PRGF in order to prevent gelatinization of the culture medium. On the other hand, mesenchymal stem cells were isolated from the umbilical cord as a primary culture. The phenotype of the cells was confirmed by flow cytometry, and the cells were randomly cultured as control (using FBS) and experimental (using PRGF) groups. The expression of the gene involved in increasing cell longevity (h-TERT) was investigated by real-time PCR technique after six days.
Results: Mesenchymal stem cells were successfully isolated from the umbilical cord. Morphologically, the mesenchymal cells cultured in the experimental group (using PRGF) were similar to the cells in the control medium. The cells exhibited a high expression level of CD73, CD90, and CD105, while the surface markers of hematopoietic cells such as CD45 and CD34 were slightly expressed. Therefore, there was no significant difference in the expression of cell surface markers between control and experimental groups. In this study, using the real-time PCR technique, it was shown that PRGF derived from the platelet could increase the expression of h-TERT gene in the umbilical cord mesenchymal stem cells compared with the control.(P = 0.034)
Conclusion: PRGF have been shown to be effective in increasing expression of h-TERT gene in the umbilical cord mesenchymal stem cells and may also be an appropriate substitute for FBS in cell culture media.
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