ABSTRACT
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
ABSTRACT
Adult stem cells [ASC] are undifferentiated cells found throughout the body. These cells are promising tools for cell replacement therapy in neurodegenerative disease. Adipose tissue is the most abundant and accessible source of ASC. This study was conducted to evaluate effect of selegiline on differentiation of adiposederived stem cells [ADSC] into functional neuron-like cells [NLC], and also level of the neurotrophin expression in differentiated cells. ADSC were transdifferentiated into NLC using selegiline where CD90, CD49d, CD31, CD106 and CD45 were used as markers for ADSC identification. Lipogenic and osteogenic differentiation of ADSC were used to characterize the ADSC. ADSC were treated with selegiline at different concentrations [from 10[-6] to 10[-11] mM] and time points [3, 6, 12, 24 and 48 h]. Percentage of viable cells, nestin and neurofilament 68 [NF-68] immunoreactive cells were used as markers for differentiation. The optimal dose for neurotrophin expressions in differentiating cells was evaluated using reverse transcriptase-PCR. NLC function was evaluated by loading and unloading with FM1-43 dye. ADSC were immunoreactive to CD90 [95.67 +/- 2.26], CD49d [71.52 +/- 6.64] and CD31 [0.6 +/- 0.86], but no immunoreactivity was detected for CD106 and CD45. The results of neural differentiation showed the highest percentage of nestin and NF-68 positive cells at 10[-9] mM concentration of selegiline [exposed for 24 h]. The differentiated cells expressed synapsin and neurotrophin genes except brainderived neurotrophic factor. ADSC can be an alternative source in cell-based therapy for neurodegenerative diseases using selegiline to induce ADSC differentiation to neuronal lineage