IBJ-Iranian Biomedical Journal. 2013; 17 (1): 8-14
在 En
| IMEMR
| ID: emr-193078
Responsible library:
EMRO
Background: 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]
Methods: BMSC isolated from the femurs and tibias of adult rats were cultured in DMEM/F12 medium supplemented with 10% FBS, pre induced using [beta]-mercaptoethanol [[beta]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
Results: 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
Conclusion: the results of this study showed that the application of [beta]ME, RA, and creatine induced the transdifferentiation of BMSC into GLNC. Iran. Biomed. J. 17 [1]: 8-14, 2013
Methods: BMSC isolated from the femurs and tibias of adult rats were cultured in DMEM/F12 medium supplemented with 10% FBS, pre induced using [beta]-mercaptoethanol [[beta]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
Results: 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
Conclusion: the results of this study showed that the application of [beta]ME, RA, and creatine induced the transdifferentiation of BMSC into GLNC. Iran. Biomed. J. 17 [1]: 8-14, 2013