title: Induction of pancreatic β cell gene expression in mesenchymal stem cells.
abstract: Transdifferentiattion potential of mesenchymal stem cells (MSCs) into insulin-producing cells (IPCs) has been suggested recently. In our recent works, we demonstrated the high performance of mouse neonate pancreas extract (MPE) in the production of functional IPCs from carcinoma stem cells. In this study, MPE was used to generate IPCs from MSCs without any genetic manipulation. To this end, bone marrow MSCs were isolated and characterized. In order to differentiate, MSCs were induced by selection of nestin-expressing cells and treatment with 100 μg/mL MPE. Morphological features of the differenti-ated cells were confirmed by dithizone staining. Immunoreactivity to insulin receptor beta, proinsulin, insulin, and C-peptide was observed by immunoflourescence. We also quantified glucose-dependent insulin production and secretion by ELISA. Real-time PCR indicated the expressions of β cell-related genes, PDX-1, INS1, INS2, EP300, and CREB1, in IPC cells. Possible pathways governed by CREB1, EP300, and PDX-1 transcription factors in differentiation of MSCs to IPCs were determined based on Gene Set Enrichment (GSE) approach at P = 0.05. Pathway discovery highlighted the negative regulatory effects of MIR124-2, HDAC5 protein, REST, and NR0B2 transcription factors on expression of CREB1, EP300, and PDX-1 and inhabitation of IPC differentiations. In contrast, a crosstalk between FOXA2 and TCF7L2 transcription factors, DNA-PK complex, KAT2B protein positively interacting with PDX-1, CREB1, EP300 resulted in the induction of IPC and following insulin production. In conclusion, we report an efficient, simple, and easy method for production of functional IPCs from MSCs by MPE treatment.
authors.affiliation: Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, P.O. Box 681499468, Khorramabad, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Mehrfarjam
authors.affiliation: Faculty of Basic Sciences, Department of Biology, University of Isfahan, P.O. Box 8174673441, Isfahan, Iran.|Research Institute of Biotechnology, Shahrekord University, P.O. Box 115, Shahrekord, Iran.
authors.forename: Fariba
authors.initials: F
authors.lastname: Esmaeili
authors.affiliation: Research Institute of Biotechnology, Shahrekord University, P.O. Box 115, Shahrekord, Iran.
authors.forename: Leila
authors.initials: L
authors.lastname: Shabani
authors.affiliation: Institute of Biotechnology, Shiraz University, Shiraz, Iran.|Division of Information Technology, Engineering & Environment, School of Information Technology and Mathematical Sciences, University of South Australia, Adelaide, Australia.|Department of Genetics and Evolution, The University of Adelaide, Adelaide, Australia.|Faculty of Science and Engineering, School of Biological Sciences, Flinders University, Adelaide, Australia.
authors.forename: Esmaeil
authors.initials: E
authors.lastname: Ebrahimie
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D002096:C-Peptide / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000378:metabolism; D007328:Insulin / Q000096:biosynthesis; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010179:Pancreas / Q000737:chemistry; D011247:Pregnancy; D014020:Tissue Extracts / Q000494:pharmacology*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D002096:C-Peptide; D007328:Insulin; D014020:Tissue Extracts; D014157:Transcription Factors; D005947:Glucose
keywords: insulin-producing cells; mesenchymal stem cells; nestin-positive cells; pancreas extract; stem cell differentiation