title: Generation of Insulin-Producing Cells From Human-Induced Pluripotent Stem Cells Using a Stepwise Differentiation Protocol Optimized With Platelet-Rich Plasma.
abstract: Studies on patient-specific human-induced pluripotent stem cells (hiPSCs) as well as a series of autologous growth factors presumably will reveal their many benefits for cell base replacement therapy in type 1 diabetes mellitus (TIDM) patients in the future. For this purpose, we established a multistep protocol by adding platelet-rich plasma (PRP) that induce the hiPSCs into insulin-producing cells (IPCs). We present here a differentiation protocol consisting of five stages in two groups including protocol with PRP and without PRP. Charac-teristics of derived IPCs in both groups were evaluated at the mRNA and protein levels, cell cycle and viability in the end stage of cell differentiation. The in vitro studies indicated the treatment of hiPSCs in the protocol with PRP resulting in differentiated cells with strong characteristics of IPCs including islet-like cells, the expression of mature and functional pancreatic beta cell specific marker genes. In addition to these pancreatic specific markers were detected by immunochemistry and Western blot. Our differentiated cells in two groups secreted insulin and C-peptide in a glucose stimulation test by ELISA showing in vitro functional. The results of the cell cycle assay confirmed that differentiation has been done. We reported for the first time that PRP might be ideal additive in the culture medium to induce pancreatic differentiation in the hiPSCs. This study provides a new approach to investigate the role of PRP in pancreatic differentiation protocols and enhance the feasibility of using patient-specific iPSCs and autologous PRP for future beta cells replacement therapies for T1DM. J. Cell. Physiol. 232: 2878-2886, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Medical Biotechnology Nanotechnology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
authors.forename: Seyed Ehsan
authors.initials: SE
authors.lastname: Enderami
authors.affiliation: Department of Medical Biotechnology Nanotechnology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.|Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran.
authors.forename: Yousef
authors.initials: Y
authors.lastname: Mortazavi
authors.affiliation: Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Department of Medical Biotechnology Nanotechnology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
authors.forename: Samad
authors.initials: S
authors.lastname: Nadri
authors.affiliation: Department of Molecular Medicine and Genetics, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Biglari
authors.affiliation: Department of Clinical Biochemistry, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
authors.forename: Reyhaneh Nassiri
authors.initials: RN
authors.lastname: Mansour
mesh_terms: D015153:Blotting, Western; D002096:C-Peptide / Q000378:metabolism; D018929:Cell Culture Techniques; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D003470:Culture Media / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D005819:Genetic Markers; D005947:Glucose / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D010641:Phenotype; D053657:Platelet-Rich Plasma / Q000378:metabolism*; D013997:Time Factors
chemical_list: D002096:C-Peptide; D003470:Culture Media; D005819:Genetic Markers; D007328:Insulin; D005947:Glucose