title: Enhanced differentiation of neural progenitor cells into neurons of the mesencephalic dopaminergic subtype on topographical patterns.
abstract: Parkinson's disease (PD) is a neurodegenerative disease attributed to the loss of midbrain dopaminergic (DA) neurons. The current lack of predictive models for this disease has been hampered by the acquirement of robust cells, posing a major barrier to drug development. Differentiation of stem cells into subtype specific cells may be guided by appropriate topographical cues but the role of topography has hitherto not been well understood. We used a Multi-Architecture (MARC) chip with various topographical structures and identified three topographies, which generate DA neurons from murine hippocampal neural progenitor cells with the highest percentage of neuronal (β-III-tubulin positive) and dopaminergic (tyrosine hydroxylase positive) populations. Analysis on single pattern structures showed that 2 μm gratings with 2 μm spacing and 2 μm height (2 μm gratings) and 2 μm gratings with hierarchical structure produced cells with the highest gene expression of TH and PITX3, with the longest neurite and highest percentage of alignment. Quantitative image analysis showed the 2 μm gratings produced cells with the highest expression of pituitary homeobox 3 (PITX3), LIM homeobox transcription factor 1 alpha (LMX1a), aldehyde dehydrogenase 1 family member A1 (ALDH1a1) and microtubule associated protein 2 (MAP2), as compared to nano-gratings and unpatterned controls. These patterns also enhance DA neuron differentiation on different substrate rigidities, as seen on both poly-dimethylsiloxane (PDMS) and tissue culture polystyrene (TCPS) substrates. These results show the use of topographical influence for neuronal subtype specification, which could be translated into a wide range of clinical applications for PD.
authors.affiliation: Mechanobiology Institute Singapore, National University of Singapore, T-Lab, #05-01, 5A Engineering Drive 1, Singapore 117411, Singapore.
authors.forename: Kenneth K B
authors.initials: KK
authors.lastname: Tan
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, EA-03-12, 9 Engineering Drive 1, Singapore 117575, Singapore.
authors.forename: Jason Y
authors.initials: JY
authors.lastname: Tann
authors.affiliation: Mechanobiology Institute Singapore, National University of Singapore, T-Lab, #05-01, 5A Engineering Drive 1, Singapore 117411, Singapore.
authors.forename: Sharvari R
authors.initials: SR
authors.lastname: Sathe
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, EA-03-12, 9 Engineering Drive 1, Singapore 117575, Singapore; Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 3 Research Link, Singapore 117602, Singapore.
authors.forename: Seok Hong
authors.initials: SH
authors.lastname: Goh
authors.affiliation: Duke-NUS Graduate Medical School Singapore, 8 College Road, Singapore 169857, Singapore.
authors.forename: Dongliang
authors.initials: D
authors.lastname: Ma
authors.affiliation: Duke-NUS Graduate Medical School Singapore, 8 College Road, Singapore 169857, Singapore.
authors.forename: Eyleen L K
authors.initials: EL
authors.lastname: Goh
authors.affiliation: Mechanobiology Institute Singapore, National University of Singapore, T-Lab, #05-01, 5A Engineering Drive 1, Singapore 117411, Singapore; Department of Biomedical Engineering, National University of Singapore, EA-03-12, 9 Engineering Drive 1, Singapore 117575, Singapore; Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, NUHS Tower Block, 1E Kent Ridge Road, Singapore 119228, Singapore. Electronic address: eyim@nus.edu.sg.
authors.forename: Evelyn K F
authors.initials: EK
authors.lastname: Yim
mesh_terms: D000080924:Aldehyde Dehydrogenase 1 Family; D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation; D019070:Cell Lineage; D002722:Chlorobenzenes / Q000737:chemistry; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004298:Dopamine / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology; D018398:Homeodomain Proteins / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D060850:LIM-Homeodomain Proteins / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry; D058953:Neural Stem Cells / Q000166:cytology*; D016501:Neurites / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D011137:Polystyrenes / Q000737:chemistry; D050697:Retinal Dehydrogenase / Q000378:metabolism; D015398:Signal Transduction; D013388:Succinimides / Q000737:chemistry; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods; D014157:Transcription Factors / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D002722:Chlorobenzenes; D004129:Dimethylpolysiloxanes; D018398:Homeodomain Proteins; D007527:Isoenzymes; D060850:LIM-Homeodomain Proteins; C489692:Lmx1a protein, mouse; D008869:Microtubule-Associated Proteins; C079240:Mtap2 protein, mouse; D011137:Polystyrenes; D013388:Succinimides; D014157:Transcription Factors; C433629:homeobox protein PITX3; C049563:N-(3,4,5-trichlorophenyl)succinimide; C013830:baysilon; D014446:Tyrosine 3-Monooxygenase; D000080924:Aldehyde Dehydrogenase 1 Family; C515028:ALDH1A1 protein, mouse; D050697:Retinal Dehydrogenase; D004298:Dopamine
keywords: Dopaminergic neuronal differentiation; Mid-brain neuronal patterning; Nanotopography; Neural induction; Neural tissue engineering; Neuronal differentiation