title: Directed Differentiation of Dopamine-Secreting Cells from Nurr1/GPX1 Expressing Murine Embryonic Stem Cells Cultured on Matrigel-Coated PCL Scaffolds.
abstract: Parkinson's disease (PD) is a progressive neurological disorder characterized by a large number of motor and non-motor features and is known as the second most common neurodegenerative disorder after Alzheimer's disease. The hallmark pathology of PD is the damage and death of dopamine-producing neurons in the substantia-nigra of midbrain. Intrastriatal transplants of fetal mesencephalon derived DAergic neurons have provided proof-of-principle for the cell replacement strategy and have demonstrated reinnervation of the denervated striatum. However, ethical, technical, and practical limitations of deploying fetal DAergic neurons as the source for cell therapy in PD have ceased the spread of this procedure into clinical practice. Embryonic stem (ES) cells have emerged as a therapeutic alternative that can proliferate extensively and generate dopamine-producing neurons. To this extent and to surmount the obstacles related to embryonic neural cells, many investigations have focused on using pluripotent stem cells for the derivation of DAergic neurons. In the present study, a mouse embryonic stem (mES) R1 cell line was generated which could stably co-express Nurr1 (an essential transcription factor in DAergic neuron development) and GPX-1 (a neuroprotective enzyme against oxidative stress). The Nurr1/GPX-1-expressing ES cells (Nurr1/GPX-1-ES) were differentiated into DAergic-like cells via a three-dimensional culture environment consisting of Poly-ε-Caprolactone (PCL) nanofibrous scaffolds embedded by Matrigel (Mtg) in the presence of specific signaling molecules. DAergic neuron-specific genes were highly expressed in ES-derived DAergic neurons cultured and differentiated on PCL/Mtg scaffolds. Reverse-phase HPLC confirmed that the Nurr1/GPX-1-ES-cells differentiated on PCL/Mtg electrospun scaffolds could efficiently and exclusively secrete dopamine in response to stimulus. In conclusion, our results demonstrated that PCL/Matrigel nanofibrous scaffolds could efficiently support and promote the generation of functional DAergic-like cells from Nurr1/GPX-1-ES cells. The results of this study may have an impact on future tissue engineering for cell therapy of PD.
authors.affiliation: Department of Cellular and Molecular Biology, School of Biology, University College of Science, University of Tehran, Tehran, Iran.
authors.forename: Panieh
authors.initials: P
authors.lastname: Terraf
authors.affiliation: Department of Tissue Engineering, Faculty of Advanced Medical Technologies, Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran, Iran. hbabaloo92@gmail.com.
authors.forename: Hamideh
authors.initials: H
authors.lastname: Babaloo
authors.affiliation: Department of Cellular and Molecular Biology, School of Biology, University College of Science, University of Tehran, Tehran, Iran.
authors.forename: Shideh Montasser
authors.initials: SM
authors.lastname: Kouhsari
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000008:administration & dosage*; D004298:Dopamine / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000187:drug effects / Q000378:metabolism*; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007797:Laminin / Q000008:administration & dosage*; D051379:Mice; D018345:Mice, Knockout; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000096:biosynthesis* / Q000235:genetics; D011091:Polyesters / Q000008:administration & dosage*; D011509:Proteoglycans / Q000008:administration & dosage*; D054457:Tissue Scaffolds; D000094765:Glutathione Peroxidase GPX1
chemical_list: D004338:Drug Combinations; D007797:Laminin; C539883:Nr4a2 protein, mouse; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D011091:Polyesters; D011509:Proteoglycans; C059485:matrigel; C016240:polycaprolactone; D003094:Collagen; D005979:Glutathione Peroxidase; D004298:Dopamine; D000094765:Glutathione Peroxidase GPX1
keywords: Dopaminergic neurons; Mouse embryonic stem cells; Nanofibrous scaffolds; Nurr1; PCL; Parkinson’s disease