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Human Umbilical Cord–Derived Mesenchymal Stem Cells Into Oligodendrocyte-Like Cells Using Triiodothyronine As an Inducer: A Rapid and Efficient Protocol Publisher



Almasiturk S1 ; Hamidabadi HG2, 3 ; Farzadinia P1 ; Movahed A4 ; Salimipour H1 ; Nemati R1 ; Roozbehi A5 ; Bojnordi MN2, 3 ; Darabi S6 ; Mojaverrostami S7
Authors
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Authors Affiliations
  1. 1. The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
  2. 2. Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
  3. 3. Immunogenetic Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
  4. 4. The Persian Gulf Tropical Medicine Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
  5. 5. Cellular and Molecular Research Center, Yasuj University of Medical Sciences, Yasuj, Iran
  6. 6. Cellular and Molecular Research Center, Qazvin University of Medical Science, Qazvin, Iran
  7. 7. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

Source: Regenerative Engineering and Translational Medicine Published:2023


Abstract

Abstract: Multiple sclerosis (MS) is a central nervous system (CNS)–specific autoimmune disease which is associated with severe destruction of myelin. One approach for rescuing MS patients is regenerating myelin-forming cells (Oligodendrocytes) by stem cell–based therapy. In the current work, human umbilical cord–derived mesenchymal stem cells (hUC-MSCs) were isolated from umbilical cords that were obtained postpartum. We used two steps of induction protocols to differentiate mesenchymal stem cells into oligodendrocyte-like cells: pre-induction stage and induction stage. For pre-induction stage, huc-MSCs were cultured in DMEM-F12 medium containing FBS 2% and 1 μM all-transretinoic acid for 2 days. For induction stage, firstly, the pre-induced cells were incubated in DMEM/F12 medium for 2 days in the presence of different growth factors such as; 5 ng/ml platelet-derived growth factor, 10 ng/ml basic fibroblast growth factor (bFGF). To this end, different triiodothyronine concentrations (T3): 0, 5, 12.5, 25, 50, 100, 200, and 400 ng/ml were added to the medium for 2 days. At the induction stage, the highest percentage of viability rate was found in low concentrations of T3 (5, 12.5 and 25 ng/ml). The immunofluorescence results indicated that expressions of Olig2 and O4 were just detected in cells treated by T3, and maximum protein expression was recorded in T25 ng/ml concentration. The maximum mRNA expression of nestin was detected in pre-induction stage; however, the expressions of Olig2 and PDGFR-α were significantly higher at the end of induction stage. Our work showed that hUC-MSCs can potentially differentiate into oligodendrocyte-like cells by using T3 as an inducer which can be used in the future as a promising therapeutic strategy in MS disease. Lay Summary: The highest percentage of viability rate was found in low concentrations of T3 (5, 12.5, and 25 ng/ml). In addition, our immunofluorescence data indicated that expressions of Olig2 and O4 (oligodendrocyte markers) were just detected in cells treated with T3, and maximum protein expression was recorded in T25 ng/ml concentration. The maximum mRNA expression of nestin was detected in the pre-induction stage. However, the expressions of Olig2 and PDGFR-α were significantly higher at the end of the induction stage. Our work showed that hUC-MSCs could potentially differentiate into oligodendrocyte-like cells by using T3 as an inducer which can be used in the future as a promising therapeutic strategy in MS disease. © 2023, The Author(s), under exclusive licence to The Regenerative Engineering Society.
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