Journal of Kerman University of Medical Sciences (Jul 2015)
Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors
Abstract
Background & Aims: Type 1 diabetes, or insulin-dependent diabetes, is an autoimmune disease in which pancreatic beta cells are destroyed by the immune system. Hitherto, no definite treatment has been found for this condition. Mesenchymal stem cells (MSCs) are multipotent, self-renewing cells that have the ability to differentiate into mesodermal tissues. This ability has attracted the attention of researchers toward MSCs as therapeutic agents. The aim of this study was to inspect the in vitro differentiation of human adipose-derived tissue stem cells (hADSCs) into insulin producing cells (IPCs) using minimal differentiation factors to provide a source of cells for the purpose of diabetic cell therapy. Methods: The hADSCs were obtained from liposuction aspirates and induced to differentiate into IPCs under a two-stage protocol. In the pre-induction stage, a combination of low-glucose DMEM medium, 20% (FBS), β-mercaptoethanol, and nicotinamide, and in the induction stage, high-glucose DMEM, β- mercaptoethanol, and nicotinamide without FBS was used. Differentiation was evaluated through morphological analysis, dithizone (DTZ) staining, and reverse transcription polymerase chain reaction (RTPCR). In order to evaluate the performance of differentiated cells, insulin production level was measured. Results: Morphological changes were observed using an inverted microscope at the end of the differentiation stage. Based on dithizone staining, differentiated cells were positive. Furthermore, RT-PCR confirmed the expression of insulin, pancreatic duodenal homeobox (PDX-1), paired box gene 4 (PAX-4), and glucose transporter type 2 (GLUT2) in differentiated cells. Moreover, insulin production by the IPCs was confirmed using enzyme-linked immunosorbent assay (ELISA). Conclusion: It can be concluded that hADSCs can differentiate into IPCs using minimal differentiation factors.
