title: Differentiation of Human Dental Pulp Stem Cells into Dopaminergic Neuron-like Cells in Vitro.
abstract: We investigated the potential of human dental pulp stem cells (hDPSCs) to differentiate into dopaminergic neurons in vitro as an autologous stem cell source for Parkinson's disease treatment. The hDPSCs were expanded in knockout-embryonic stem cell (KO-ES) medium containing leukemia inhibitory factor (LIF) on gelatin-coated plates for 3-4 days. Then, the medium was replaced with KO-ES medium without LIF to allow the formation of the neurosphere for 4 days. The neurosphere was transferred into ITS medium, containing ITS (human insulin-transferrin-sodium) and fibronectin, to select for Nestin-positive cells for 6-8 days. The cells were then cultured in N-2 medium containing basic fibroblast growth factor (FGF), FGF-8b, sonic hedgehog-N, and ascorbic acid on poly-l-ornithine/fibronectin-coated plates to expand the Nestin-positive cells for up to 2 weeks. Finally, the cells were transferred into N-2/ascorbic acid medium to allow for their differentiation into dopaminergic neurons for 10-15 days. The differentiation stages were confirmed by morphological, immunocytochemical, flow cytometric, real-time PCR, and ELISA analyses. The expressions of mesenchymal stem cell markers were observed at the early stages. The expressions of early neuronal markers were maintained throughout the differentiation stages. The mature neural markers showed increased expression from stage 3 onwards. The percentage of cells positive for tyrosine hydroxylase was 14.49%, and the amount was 0.526 ± 0.033 ng/mL at the last stage. hDPSCs can differentiate into dopaminergic neural cells under experimental cell differentiation conditions, showing potential as an autologous cell source for the treatment of Parkinson's disease.
authors.affiliation: BioMedical Research Institute, Kyungpook National University Hospital, Daegu, Korea.
authors.forename: So Young
authors.identifier: 0000-0003-4500-4956
authors.initials: SY
authors.lastname: Chun
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, NC, USA.
authors.forename: Shay
authors.identifier: 0000-0002-8458-9232
authors.initials: S
authors.lastname: Soker
authors.affiliation: Department of Neural Development and Disease, Korea Brain Research Institute, Daegu, Korea.
authors.forename: Yu-Jin
authors.identifier: 0000-0001-5660-3001
authors.initials: YJ
authors.lastname: Jang
authors.affiliation: Department of Urology, Kyungpook National University, Daegu, Korea.
authors.forename: Tae Gyun
authors.identifier: 0000-0002-4390-0952
authors.initials: TG
authors.lastname: Kwon
authors.affiliation: Department of Urology, Kyungpook National University, Daegu, Korea.
authors.forename: Eun Sang
authors.identifier: 0000-0002-7442-6886
authors.initials: ES
authors.lastname: Yoo
mesh_terms: D000818:Animals; D001921:Brain / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D003782:Dental Pulp / Q000166:cytology*; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000378:metabolism* / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D004676:Myelin Basic Protein / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D055746:Stage-Specific Embryonic Antigens / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism* / Q000473:pathology; D014404:Tubulin / Q000235:genetics / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis / Q000235:genetics / Q000378:metabolism
chemical_list: D003470:Culture Media; D005904:Glial Fibrillary Acidic Protein; D004676:Myelin Basic Protein; D055746:Stage-Specific Embryonic Antigens; C498585:TUBB3 protein, human; D014404:Tubulin; C066584:stage-specific embryonic antigen-4; D014446:Tyrosine 3-Monooxygenase
keywords: Autologous Stem Cell Therapy; Dopaminergic Differentiation; Dopaminergic Neural Cells; Human Dental Pulp Stem Cells; Parkinson Disease; Tyrosine Hydroxylase
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