title: miR-375 induces human decidua basalis-derived stromal cells to become insulin-producing cells.
abstract: This paper focuses on the development of renewable sources of isletreplacement tissue for the treatment of type I diabetes mellitus. Placental tissue-derived mesenchymal stem cells (MSCs) are a promising source for regenerative medicine due to their plasticity and easy availability. They have the potential to differentiate into insulin-producing cells. miR-375 is a micro RNA that is expressed in the pancreas and involved in islet development. Human placental decidua basalis MSCs (PDB-MSCs) were cultured from full-term human placenta. The immunophenotype of the isolated cells was checked for CD90, CD105, CD44, CD133 and CD34 markers. The MSCs (P3) were chemically transfected with hsa-miR-375. Total RNA was extracted 4 and 6 days after transfection. The expressions of insulin, NGN3, GLUT2, PAX4, PAX6, KIR6.2, NKX6.1, PDX1, and glucagon genes were evaluated using real-time qPCR. On day 6, we tested the potency of the clusters in response to the high glucose challenge and assessed the presence of insulin and NGN3 proteins via immunocytochemistry. Flow cytometry analysis confirmed that more than 90% of the cells were positive for CD90, CD105 and CD44 and negative for CD133 and CD34. Morphological changes were followed from day 2. Cell clusters formed during day 6. Insulin-producing clusters showed a deep red color with DTZ. The expression of pancreatic-specific transcription factors increased remarkably during the four days after transfection and significantly increased on day 7. The clusters were positive for insulin and NGN3 proteins, and C-peptide and insulin secretion increased in response to changes in the glucose concentration (2.8 mM and 16.7 mM). In conclusion, the MSCs could be programmed into functional insulin-producing cells by transfection of miR-375.
authors.affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Fars, Iran.
authors.forename: Anahita
authors.initials: A
authors.lastname: Shaer
authors.forename: Negar
authors.initials: N
authors.lastname: Azarpira
authors.forename: Akbar
authors.initials: A
authors.lastname: Vahdati
authors.forename: Mohammad Hosein
authors.initials: MH
authors.lastname: Karimi
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Shariati
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D003656:Decidua / Q000166:cytology*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D005947:Glucose / Q000494:pharmacology; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D010920:Placenta / Q000166:cytology; D011247:Pregnancy; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D015703:Antigens, CD; D051792:Basic Helix-Loop-Helix Proteins; D005947:Glucose; D018398:Homeodomain Proteins; D007328:Insulin; D035683:MicroRNAs; D009419:Nerve Tissue Proteins; D015534:Trans-Activators; C540933:MIRN375 microRNA, human; C498695:NEUROG3 protein, human; C084010:pancreatic and duodenal homeobox 1 protein