title: MicroRNA-145-based differentiation of human mesenchymal stem cells to smooth muscle cells.
abstract: OBJECTIVE
  To investigate the role of microRNA-145, that regulates gene expression of genes related to differentiation, proliferation and the phenotype of smooth muscle cells (SMCs), in the differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) to SMCs.
  
  
  RESULTS
  Real-time PCR analysis indicated significant upregulation of SMC markers, including SM-α-actin, calponin, caldesmon and SMMHC, in SMCs compared to hBM-MSCs. Conversely, Krüppel-like factor 4, the direct target of microRNA-145 and the suppressor of smooth muscle differentiation, was suppressed in hBM-MSC-derived SMCs. Western blot analysis and immunocytochemistry also confirmed that the introduction of microRNA-145 into hBM-MSCs induced mature contractile SMCs. The functionality of hBM-MSC-derived SMCs was assessed by proliferation assay using PDGF-BB and contractility assay using carbachol. The results showed that the produced SMCs contracted in response to carbachol stimulation.
  
  
  CONCLUSIONS
  Overexpression of microRNA-145 in undifferentiated hBM-MSCs results in functionally mature contractile SMCs that can be used in drug discovery and cell therapy in SMC disorders such as vascular disease.
authors.affiliation: Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.|Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Pajoohesh
authors.affiliation: Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran. naderman@modares.ac.ir.
authors.forename: Hossein
authors.initials: H
authors.lastname: Naderi-Manesh
authors.affiliation: Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
mesh_terms: D000199:Actins / Q000235:genetics; D002135:Calcium-Binding Proteins / Q000235:genetics; D002148:Calmodulin-Binding Proteins / Q000235:genetics; D002217:Carbachol / Q000494:pharmacology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D035683:MicroRNAs / Q000235:genetics*; D008840:Microfilament Proteins / Q000235:genetics; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000187:drug effects; D000096985:Calponins
chemical_list: D000199:Actins; D002135:Calcium-Binding Proteins; D002148:Calmodulin-Binding Proteins; D002217:Carbachol; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D035683:MicroRNAs; D008840:Microfilament Proteins; D000096985:Calponins; C000715467:KLF4 protein, human; C541010:MIRN145 microRNA, human
keywords: Bone marrow mesenchymal stem cell; Differentiation; Krüppel-like factor 4; MicroRNA-145; Smooth muscle cell