title: In vitro reprogramming of rat bmMSCs into pancreatic endocrine-like cells.
abstract: Islet transplantation provides curative treatments to patients with type 1 diabetes, but donor shortage restricts the broad use of this therapy. Thus, generation of alternative transplantable cell sources is intensively investigated worldwide. We previously showed that bone marrow-derived mesenchymal stem cells (bmMSCs) can be reprogrammed to pancreatic-like cells through simultaneously forced suppression of Rest/Nrsf (repressor element-1 silencing transcription factor/neuronal restrictive silencing factor) and Shh (sonic hedgehog) and activation of Pdx1 (pancreas and duodenal transcription factor 1). We here aimed to reprogram bmMSCs further along the developmental pathway towards the islet lineages by improving our previous strategy and by overexpression of Ngn3 (neurogenin 3) and NeuroD1 (neurogenic differentiation 1), critical regulators of the development of endocrine pancreas. We showed that compared to the previous protocol, the overexpression of only Pdx1 and Ngn3 reprogrammed bmMSCs into cells with more characteristics of islet endocrine lineages verified with bioinformatic analyses of our RNA-Seq datasets. These analyses indicated 2325 differentially expressed genes including those involved in the pancreas and islet development. We validated with qRT-PCR analysis selective genes identified from the RNA-Seq datasets. Thus, we reprogrammed bmMSCs into islet endocrine-like cells and advanced the endeavor to generate surrogate functional insulin-secreting cells.
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology of Ministry of Public Health, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, 110013, China.|Key Laboratory of Genetics and Reproduction Health of National Health and Family Planning Commission, Liaoning Province Research Institute of Family Planning, Shenyang, 110031, China.
authors.forename: Hong-Tu
authors.initials: HT
authors.lastname: Li
authors.affiliation: Islet Cell Development Program, Harry Perkins Institute of Medical Research, and Centre for Medical Research, 6 Verdun St, Nedlands, WA, Australia. fangxu.jiang@perkins.uwa.edu.au.|University of Western Australia, Perth, 6009, Australia. fangxu.jiang@perkins.uwa.edu.au.
authors.forename: Fang-Xu
authors.initials: FX
authors.lastname: Jiang
authors.affiliation: Department of General Practice, The First Affiliated Hospital, China Medical University, Shenyang, 110001, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Shi
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology of Ministry of Public Health, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, 110013, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology of Ministry of Public Health, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, 110013, China.
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: Liu
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology of Ministry of Public Health, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, 110013, China.
authors.forename: Xue-Wen
authors.initials: XW
authors.lastname: Lin
authors.affiliation: National key laboratory of endocrinology, China Medical University, Shenyang, 110001, China.
authors.forename: Zhong-Yan
authors.initials: ZY
authors.lastname: San
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology of Ministry of Public Health, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, 110013, China. pangxining@126.com.
authors.forename: Xi-Ning
authors.initials: XN
authors.lastname: Pang
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D065150:Cellular Reprogramming; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D063990:Gene Ontology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D017423:Sequence Analysis, RNA; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection
chemical_list: D007328:Insulin; D014157:Transcription Factors
keywords: Lentivirus vector; Mesenchymal stem cells; Pancreatic endocrine lineage cells; RNA-Seq; Reprogramming