title: Exogenous Expression of Nt-3 and TrkC Genes in Bone Marrow Stromal Cells Elevated the Survival Rate of the Cells in the Course of Neural Differentiation.
abstract: Bone marrow stromal cells (BMSCs) are attractive cellular sources for cell therapy of many diseases, specifically neurodegenerative ones. The potential capability of BMSCs could be further augmented by enhancing their neuroprotective property, differentiation potential, and survival rate subsequent to transplantation. Therefore, a concurrent upregulation of neurotrophin-3 (NT-3) and its high affinity receptor, tyrosin kinase C (TrkC), was utilized in our study. BMSCs were cotransfected with pDsRed1-N1-NT-3 and pCMX-TrkC plasmids before induction of neural differentiation. pEGFP-N1-transfected BMSCs were also employed as a control. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed for gene expression analysis. Cell viability was evaluated by MTT assay, while apoptosis rate was assessed by flow cytometry after PI and Annexin V staining. NT-3 and TrkC mRNA levels were greatly elevated following cotransfection of cells with pDsRed1-N1-NT-3 and pCMX-TrkC vectors. The expression of neural markers (i.e., NFM, and NeuroD1) was augmented in cotransfected BMSCs, compared to the control ones, after neural induction. At each time point, the viability and apoptosis rates of the cells over-expressing NT-3 and TrkC showed increased and reduced patterns, respectively. Our data demonstrated that NT-3/TrkC-co-transfected BMSCs, compared to those of intact cells, could be more beneficial graft candidates for the upcoming treatment strategies of neurogenic disorders due to their increased viability and expression of neural markers. This may be due to their increased level of neural differentiation potential and/or their enhanced rate of survival and/or their useful capacity to secrete NT-3.
authors.affiliation: Genetic Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Houri
authors.initials: H
authors.lastname: Edalat
authors.affiliation: Department of Physiology, Aerospace Research Institute, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Hajebrahimi
authors.affiliation: Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Vahid
authors.initials: V
authors.lastname: Pirhajati
authors.affiliation: Genetic Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Tavallaei
authors.affiliation: Department of Anatomy, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mansoureh
authors.initials: M
authors.lastname: Movahedin
authors.affiliation: Department of Molecular Genetics, School of Biological Sciences, Tarbiat Modares University, Tehran, Iran. sjmowla@modares.ac.ir.
authors.forename: Seyed Javad
authors.initials: SJ
authors.lastname: Mowla
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D015870:Gene Expression; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D020933:Neurotrophin 3 / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020812:Receptor, trkC / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D020933:Neurotrophin 3; D020812:Receptor, trkC
keywords: Apoptosis; Bone marrow stromal cells; Cell viability; NT-3; Neural differentiation; TrkC