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The Potency of Hsa-Mir-9-1 Overexpression in Photoreceptor Differentiation of Conjunctiva Mesenchymal Stem Cells on a 3D Nanofibrous Scaffold Publisher Pubmed



Rahmani A1 ; Naderi M2 ; Barati G3 ; Arefian E4 ; Jedari B3 ; Nadri S1, 5, 6, 7
Authors
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Authors Affiliations
  1. 1. Department of Medical Nanotechnology, Zanjan University of Medical Sciences, Zanjan, Iran
  2. 2. Cell-Based Therapies Research Center, Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Department of Medical Biotechnology, Zanjan University of Medical Sciences, Zanjan, Iran
  4. 4. Department of Microbiology, School of Biology, College of Science, University of Tehran, Iran
  5. 5. Zanjan Metabolic Diseases Research Center, Zanjan University of Medical Sciences, Zanjan, Iran
  6. 6. Zanjan Pharmaceutical Nanotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran
  7. 7. Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran

Source: Biochemical and Biophysical Research Communications Published:2020


Abstract

MiRNAs are small non-coding RNAs that are ordinarily involved in modulating mRNAs and stem cell differentiation. 3D nanofibrous scaffolds have an important role in the differentiation of stem cells due to their similarity to the extracellular matrix (ECM). In the present study, we tried to introduce a new approach to guiding the differentiation of conjunctiva mesenchymal stem cells (CJMSCs) into photoreceptor-like cells by hsa-miR-9-1 delivery on both 2D and 3D substrates. First, the CJMSCs were transduced by a lentiviral vector carrying miR-9 (pCDH + hsa-miR-9-1) and then cell transduction efficacy verified by using fluorescent microscopy, flow cytometry, and qPCR analyses. Silk Fibroin-poly-L-lactic acid (SF-PLLA) scaffold was fabricated by the electrospinning technique while the scaffold characteristics including morphology, chemical properties, and biocompatibility were evaluated by SEM, FTIR, and MTT assays, respectively. Then, the miR-9-CJMSCs were seeded on both TCPS and the scaffold; photoreceptor gene and protein expressions were evaluated by RT-qPCR and immunostaining after 14 and 21 days of transduction. More than 80% of CJMSCs were transduced and miR-9 expression was significantly higher in miR-9-CJMSCs compared with empty vector (EV)-CJMSCs. SEM and FTIR confirmed the fabrication of the SF/PLLA hybrid structure. RT-qPCR and immunostaining analyses showed that the specific photoreceptor genes and proteins were expressed in miR-9 transduced CJMSCs. Mir-9 induced CJMSCs into photoreceptor-like cells in a time-dependent manneron on both TCPS and nanofibrous scaffold.We have proved that hsa-miR-9-1 has the potency to guide the photoreceptor differentiation of mesenchymal stem cells and promote retinal regeneration. © 2020 Elsevier Inc.
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1. Synergistic Effect of Mir-9 Overexpression and Electrical Induction on Differentiation of Conjunctiva Mesenchymal Stem Cells Into Photoreceptor-Like Cells, International Journal of Artificial Organs (2022)
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