title: A new multistep induction protocol for the transdifferentiation of bone marrow stromal stem cells into GABAergic neuron-like cells.
abstract: Bone marrow stromal stem cells (BMSC) are appropriate source of multipotent stem cells that are ideally suited for use in various cell-based therapies. It can be differentiated into neuronal-like cells under appropriate conditions. This study examined the effectiveness of co-stimulation of creatine and retinoic acid in increasing the differentiation of BMSC into GABAergic neuron-like cells (GNLC).
  
  
  
  BMSC isolated from the femurs and tibias of adult rats were cultured in DMEM/F12 medium supplemented with 10% FBS, pre-induced using β-mercaptoethanol β-ME) and induced using retinoic acid (RA) and creatine. Immunostaining of neurofilament 200 kDa, neurofilament 160 kDa, nestin, fibronectin, Gamma-amino butyric acid (GABA) and glutamic acid decarboxylase (GAD) 65/67 were used to evaluate the transdifferentiation of BMSC into GNLC and to evaluate the effectiveness of pre-induction and induction assays. The expression of genes that encode fibronectin, octamer-binding transcription factor 4 (Oct-4), GAD 65/67 and the vesicular GABA transporter was examined in BMSC and GNLC by using RT-PCR assays during transdifferentiation of BMSC into GLNC.
  
  
  
  Co-stimulation with RA and creatine during the induction stage doubled the rates of GABAergic differentiation compared with induction using creatine alone, resulting in a 71.6% yield for GLNC. RT-PCR showed no expression of Oct-4 and fibronectin after the induction stage.
  
  
  
  The results of this study showed that the application of β-ME, RA, and creatine induced the transdifferentiation of BMSC into GLNC.
authors.affiliation: Dept. of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Shahram
authors.initials: S
authors.lastname: Darabi
authors.affiliation: Dept. of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.|Shefa Neurosciences Research Center, Khatam Al-Anbia Hospital,Tehran, Iran.
authors.forename: Taki
authors.initials: T
authors.lastname: Tiraihi
authors.affiliation: Faculty of Medical Sciences, Shahed University, Tehran, Iran.
authors.forename: AliReza
authors.initials: A
authors.lastname: Delshad
authors.affiliation: Dept. of Genetics, Faculty of Basic Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Sadeghizadeh
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D054338:Cell Transdifferentiation / Q000187:drug effects*; D002478:Cells, Cultured; D003401:Creatine; D005260:Female; D005353:Fibronectins / Q000096:biosynthesis; D059330:GABAergic Neurons / Q000166:cytology / Q000378:metabolism*; D005968:Glutamate Decarboxylase / Q000096:biosynthesis; D008623:Mercaptoethanol; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014212:Tretinoin; D050495:Vesicular Inhibitory Amino Acid Transport Proteins / Q000096:biosynthesis
chemical_list: D005353:Fibronectins; D050814:Octamer Transcription Factor-3; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; C493406:vesicular GABA transporter; D014212:Tretinoin; D008623:Mercaptoethanol; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2; D003401:Creatine