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Nowadays, cell replacement using stem cells has been considered as an important potential therapy for treatment of Parkinson’s disease. Since the production of functional dopaminergic neurons is the first step in cell-replacement therapy, herein, we aimed to develop a high efficiency method to generate dopaminergic neurons from adipose derived stem cells (ADSCs) obtained from the adult adipose tissue. ADSCs were isolated and treated with growth factors for 2 weeks. Next, immunoreactivity for neuronal markers nestin, 2-tubulin3, and microtubule-associated protein 2 and dopaminergic specific marker tyrosin hydroxylase was evaluated. Moreover, mRNA expression nestin, MAP2, PITX3, TH and KCNH5 was measured using quantitative real time PCR. Electrophysiological properties and ability of dopamine secretion were evaluated using current clamp and HPLC methods Our data. revealed that ADSCs showed the morphological changes following neuronal induction after 14 weeks. Furthermore, ADSCs-derived dopaminergic neurons were immunopositive for neuronal and dopaminergic-specific markers. In addition, mRNA expression of nestin, MAP2, PITX3, TH and KCNH5 significantly increased in ADSCs-derived dopaminergic neurons. Finally, electrophysiological results confirmed that these neurons are able to excitation and they could produce action potential. Moreover, interestingly, HPLC analysis indicated that this neuronal cells have capacity of dopamine production and secretion that was in line with over-expression of TH enzyme in these cells. although we provided a collection of data about the functional dopaminergic neurons derived from ADSCs, further in vivo studies are required to confirm the suitability of these dopaminergic cells for cell replacement in disorder related to dopaminergic neurons lesions.

In vitro Differentiation of Adipose Derived Stem Cells into Functional Dopaminergic Neurons