期刊
STEM CELLS TRANSLATIONAL MEDICINE
卷 5, 期 11, 页码 1525-1537出版社
WILEY
DOI: 10.5966/sctm.2015-0318
关键词
Diabetes; Insulin-producing cells; SCID-beige mice; Synthetic modified mRNA; V-Maf musculoaponeurotic fibrosarcoma oncogene homolog A (MAFA)
资金
- Belgian Society for Pediatric Endocrinology and Diabetology (BESPEED)
- Institut de Recherche Clinique et Experimentale (IREC)
- Fonds National de la Recherche Scientifique (FNRS)
- International Society for Pediatric and Adolescent Diabetes (ISPAD)
beta-Cell replacement therapy represents the most promising approach to restore beta-cell mass and glucose homeostasis in patients with type 1 diabetes. Safety and ethical issues associated with pluripotent stem cells stimulated the search for adult progenitor cells with endocrine differentiation capacities. We have already described a model for expansion and differentiation of human pancreatic duct-derived cells (HDDCs) into insulin-producing cells. Here we show an innovative and robust in vitro system for large-scale production of beta-like cells from HDDCs using a nonintegrative RNA-based reprogramming technique. Synthetic modified RNAs for pancreatic transcription factors (pancreatic duodenal homeobox 1, neurogenin3, and V-Maf musculoaponeurotic fibrosarcoma oncogene homolog A [MAFA]) were manufactured and daily transfected in HDDCs without strongly affecting immune response and cell viability. MAFA overexpression was efficient and sufficient to induce beta-cell differentiation of HDDCs, which acquired a broad repertoire of mature beta-cell markers while downregulating characteristic epithelial-mesenchymal transition markers. Within 7 days, MAFA-reprogrammed HDDC populations contained 37% insulin-positive cells and a proportion of endocrine cells expressing somatostatin and pancreatic polypeptide. Ultrastructure analysis of differentiated HDDCs showed both immature and mature insulin granules with light-backscattering properties. Furthermore, in vitro HDDC-derived beta cells (called beta-HDDCs) secreted human insulin and C-peptide in response to glucose, KCI, 3-isobutyl-1-methylxanthine, and tolbutamide stimulation. Transplantation of beta-HDDCs into diabetic SCID-beige mice confirmed their functional glucose-responsive insulin secretion and their capacity to mitigate hyperglycemia. Our data describe a new, reliable, and fast procedure in adult human pancreatic cells to generate clinically relevant amounts of new beta cells with potential to reverse diabetes.
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