In Vitro Differentiation of Adipose Derived Stem Cells Into Functional Dopaminergic Neurons
Azam Asemi Rad1, Mohammad Hasan Heidari1, Abbas Aliaghaei1, Mehdi Eskandarian Broujeni3, Amir Shojaei4, Hojjat-Allah Abbaszadeh2, Fatemeh Shaerzadeh5 and Yousef Sadeghi21Anatomy and cell biology department, School of medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
2Hearing disorders research center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
3Stem cells and regenerative medicine, Faculty of medical biotechnology, National institute of genetic engineering and biotechnology, Tehran, Iran.
4Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
5Molecular Medicine Research Center, Hormozgan University of Medical Sciences, Bandar Abbas, Iran; Department of Physiology, faculty of medicine, Hormozgan University of Medical Sciences, Bandar Abbas, Iran.
Corresponding Author E-mail: dr.ysadeghi@yahoo.com
Abstract: 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, β-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.
Keywords: dopamine Parkinson disease; neural Stem cell; tyrosine hydroxylase; Back to TOC