In vitro generation of functional dopaminergic neurons from full-term amniotic fluid stem cells

Abstract

Event Abstract Back to Event In vitro generation of functional dopaminergic neurons from full-term amniotic fluid stem cells Siti Nurussaadah Hamzah1, 2, Hoo Mun-Fun1, 2, Sharmili Vidyadaran3 and Norshariza Nordin1, 2* 1 Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, Genetics and Regenerative Medicine Research Centre, Malaysia 2 Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, Department of Biomedical Science, Malaysia 3 Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, Department of Pathology, Malaysia Stem cell replacement therapy has been seen as a promising alternative treatment for Parkinson’s disease. Unfortunately, the efficiency in generating functional dopaminergic (DA) neurons from stem cells remains a great challenge. Different stem cell types have different dopaminergic potential that require different differentiation protocols. Amniotic fluid stem cells (AFSCs) are among the prominent stem cell sources in regenerative medicine that have broad differentiation capacity with less safety and ethical concerns. Here, we aimed to explore the differentiation potential of our newly established full-term rat AFSCs in generating DA neurons and evaluate their functional properties and maturity in-vitro. Two methods; adherent monoculture and multicellular aggregation (embryoid bodies, EBs) with addition of all-trans retinoic acid (4-/4+) protocols, were used to differentiate the AFSCs into neurons. Both methods clearly showed AFSCs to be highly neurogenic and successfully generated cells with good neuron-like morphology expressing neural precursor marker (Nestin), post-mitotic and mature neuronal markers (Class III β-tubulin and Microtubule-associated protein 2; MAP2) and DA marker (Tyrosine hydroxylase; TH). We further examined the effects of patterning factors; fibroblast growth factor 8b (Fgf8b) and sonic hedgehog (Shh) using the adherent monoculture method in generating DA neurons. The differentiated cells expressed DA neuronal markers MAP2 and TH, midbrain DA marker (Pituitary homeobox 3; Pitx3), neuronal maturity (G-protein-regulated inward-rectifier potassium (K+) channel 2; GIRK2) and functionality markers (Vesicular monoamine transporter 2; VMAT2 and Synaptophysin). Interestingly, no significant difference was observed in expression of these markers by the cells generated with and without the addition of Fgf8b and Shh. The neurons were also observed to release dopamine in the absence of these factors. These results highly suggest that full-term AFSCs as a highly potential source for stem cell-derived DA neurons thus offer as a valuable tool in the endeavour towards PD therapy. Acknowledgements This project was supported by Research University Grant Scheme (RUGS): project number 04-02-12-1758RU, from Universiti Putra Malaysia awarded to N Nordin, and Special Graduate Research Award (SGRA) from Universiti Putra Malaysia awarded to SN Hamzah. Keywords: Amniotic Fluid, Dopamine, Stem Cells, in vitro, neurone Conference: 14th Meeting of the Asian-Pacific Society for Neurochemistry, Kuala Lumpur, Malaysia, 27 Aug - 30 Aug, 2016. Presentation Type: Poster Presentation Session Topic: 14th Meeting of the Asian-Pacific Society for Neurochemistry Citation: Hamzah S, Mun-Fun H, Vidyadaran S and Nordin N (2016). In vitro generation of functional dopaminergic neurons from full-term amniotic fluid stem cells. Conference Abstract: 14th Meeting of the Asian-Pacific Society for Neurochemistry. doi: 10.3389/conf.fncel.2016.36.00185 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. 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Received: 04 Aug 2016; Published Online: 11 Aug 2016. * Correspondence: Dr. Norshariza Nordin, Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, Genetics and Regenerative Medicine Research Centre, Serdang, Selangor, Malaysia, shariza@upm.edu.my Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Siti Nurussaadah Hamzah Hoo Mun-Fun Sharmili Vidyadaran Norshariza Nordin Google Siti Nurussaadah Hamzah Hoo Mun-Fun Sharmili Vidyadaran Norshariza Nordin Google Scholar Siti Nurussaadah Hamzah Hoo Mun-Fun Sharmili Vidyadaran Norshariza Nordin PubMed Siti Nurussaadah Hamzah Hoo Mun-Fun Sharmili Vidyadaran Norshariza Nordin Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.