Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors
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 tha...
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| Format: | Article |
| Language: | English |
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Kerman University of Medical Sciences
2015-07-01
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| Series: | Journal of Kerman University of Medical Sciences |
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| Online Access: | https://jkmu.kmu.ac.ir/article_16085_33ba8bff40a2bc26ffd69b73fa920a2f.pdf |
| _version_ | 1797790695751155712 |
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| author | Iman Rad Alireza Farsinejad Mohammad-Amin Edalatmanesh |
| author_facet | Iman Rad Alireza Farsinejad Mohammad-Amin Edalatmanesh |
| author_sort | Iman Rad |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-13T02:08:13Z |
| format | Article |
| id | doaj.art-7fa5ae9c758147c6926360b1c534cdad |
| institution | Directory Open Access Journal |
| issn | 2008-2843 |
| language | English |
| last_indexed | 2024-03-13T02:08:13Z |
| publishDate | 2015-07-01 |
| publisher | Kerman University of Medical Sciences |
| record_format | Article |
| series | Journal of Kerman University of Medical Sciences |
| spelling | doaj.art-7fa5ae9c758147c6926360b1c534cdad2023-07-01T05:24:30ZengKerman University of Medical SciencesJournal of Kerman University of Medical Sciences2008-28432015-07-0122432834016085Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation FactorsIman Rad0Alireza Farsinejad1Mohammad-Amin Edalatmanesh2Physiology Research Center, Kerman University of Medical Sciences, Kerman, IranAssistant Professor of Pathology, Stem Cell Research Center, Kerman University of Medical Sciences, Kerman, IranAssistant Professor, Department of Physiology, College of Sciences, Shiraz Branch, Islamic Azad University, Shiraz, IranBackground & 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.https://jkmu.kmu.ac.ir/article_16085_33ba8bff40a2bc26ffd69b73fa920a2f.pdfdiabetesmesenchymal stem cells (mscs)adipose tissueinsulin |
| spellingShingle | Iman Rad Alireza Farsinejad Mohammad-Amin Edalatmanesh Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors Journal of Kerman University of Medical Sciences diabetes mesenchymal stem cells (mscs) adipose tissue insulin |
| title | Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors |
| title_full | Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors |
| title_fullStr | Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors |
| title_full_unstemmed | Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors |
| title_short | Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Insulin Producing Cells Using Minimal Differentiation Factors |
| title_sort | differentiation of human adipose tissue derived mesenchymal stem cells into insulin producing cells using minimal differentiation factors |
| topic | diabetes mesenchymal stem cells (mscs) adipose tissue insulin |
| url | https://jkmu.kmu.ac.ir/article_16085_33ba8bff40a2bc26ffd69b73fa920a2f.pdf |
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