Differentiation of human adipose-derived mesenchymal stem cell into insulin-producing cells: an in vitro study
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ORIGINAL PAPER
Differentiation of human adipose-derived mesenchymal stem cell into insulin-producing cells: an in vitro study P. Rahnamay Moshtagh & S. Hojati Emami & Ali M. Sharifi
Received: 21 July 2012 / Accepted: 11 December 2012 # University of Navarra 2012
Abstract Stem cells with the ability to differentiate into insulin-producing cells (IPCs) are becoming the most promising therapy for diabetes mellitus and reduce the major limitations of availability and allogeneic rejection of beta cell transplantations. Mesenchymal stem cells (MSCs) are pluripotent stromal cells with the ability to proliferate and differentiate into a variety of cell types including endocrine cells of the pancreas. This study sought to inspect the in vitro differentiation of human adipose-derived tissue stem cells into IPCs which could provide an abundant source of cells for the purpose of diabetic cell therapy in addition to avoid immunological rejection. Adipose-derived MSCs were obtained from liposuction aspirates and induced to differentiate into insulin-secreting cells under a three-stage protocol based on a combination of low-glucose DMEM medium, β-
mercaptoethanol, and nicotinamide for pre-induction and high-glucose DMEM, β-mercaptoethanol, nicotinamide, and exendin-4 for induction stages of differentiation. Differentiation was evaluated by the analysis of morphology, dithizone staining, RT-PCR, and immunocytochemistry. Morphological changes including typical islet-like cell clusters were observed by phase-contrast microscope at the end of differentiation protocol. Based on dithizone staining, differentiated cells were positive and undifferentiated cells were not stained. Furthermore, RT-PCR results confirmed the expression of insulin, PDX1, Ngn3, PAX4, and GLUT2 in differentiated cells. Moreover, insulin production by the IPCs was confirmed by immunocytochemistry analysis. It is concluded that adipose-derived MSCs could differentiate into insulin-producing cells in vitro.
P. R. Moshtagh : A. M. Sharifi Razi Drug Research Center and Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
A. M. Sharifi Endocrine and Metabolism research Institute, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran
A. M. Sharifi Department of Tissue Engineering and cell therapy, School of advanced sciences in Medicine, Tehran University of Medical Sciences, Tehran, Iran P. R. Moshtagh : S. H. Emami Department of Biomedical Engineering, Amirkabir University of Technology, Tehran, Iran
A. M. Sharifi Cellular and Molecular Research Center, Tehran University of Medical Sciences, Tehran, Iran A. M. Sharifi (*) Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran e-mail: [email protected] A. M. Sharifi e-mail: [email protected]
P.R. Moshtagh et al.
Keywords Mesenchymal stem cells . Insulinproducing cells . Differentiation . Diabetes . Cell therapy
Introduction Diabetes is a global, complicated metabolic disease and one of the leadi
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