Academic Journal
Distinct media requirements for dopaminergic neuron reprogramming from human glia and fibroblasts.
| Τίτλος: | Distinct media requirements for dopaminergic neuron reprogramming from human glia and fibroblasts. |
|---|---|
| Συγγραφείς: | Laurin K; Developmental and Regenerative Neurobiology, Lund Stem Cell Center, Department of Experimental Medical Science, Faculty of Medicine, Lund University, Lund, Sweden., Kajtez J; Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark., Wickham J; Laboratory of Stem Cells and Restorative Neurology, Lund Stem Cell Center, Faculty of Medicine, Lund University, Lund, Sweden., Parmar M; Developmental and Regenerative Neurobiology, Lund Stem Cell Center, Department of Experimental Medical Science, Faculty of Medicine, Lund University, Lund, Sweden., Habekost M; Developmental and Regenerative Neurobiology, Lund Stem Cell Center, Department of Experimental Medical Science, Faculty of Medicine, Lund University, Lund, Sweden. mette.habekost@med.lu.se.; Center for Translational Neuromedicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, 2200, Denmark. mette.habekost@med.lu.se.; Department of Biotechnology and Biomedicine, DTU Bioengineering, Technical University of Denmark, Kgs. Lyngby, 2800, Denmark. mette.habekost@med.lu.se. |
| Πηγή: | Scientific reports [Sci Rep] 2026 Aug 17; Vol. 16 (1). Date of Electronic Publication: 2026 Aug 17. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: London : Nature Publishing Group, copyright 2011- |
| Ιατρικοί όροι (MeSH): | Dopaminergic Neurons*/cytology , Dopaminergic Neurons*/metabolism , Dopaminergic Neurons*/drug effects , Fibroblasts*/cytology , Fibroblasts*/metabolism , Fibroblasts*/drug effects , Cellular Reprogramming*/drug effects , Neuroglia*/cytology , Neuroglia*/metabolism , Neuroglia*/drug effects , Culture Media*/pharmacology , Culture Media*/chemistry, Tyrosine 3-Monooxygenase/metabolism ; Dopamine/metabolism ; Humans ; Cells, Cultured ; Cellular Reprogramming Techniques |
| Περίληψη: | Direct reprogramming of somatic cells into induced dopaminergic neurons (iDANs) is a promising strategy for replacing degenerated dopamine neurons in Parkinson's disease. One current bottleneck for translation of direct neuronal reprogramming is the low reprogramming efficiency and difficulty to acquire enough tyrosine hydroxylase (TH) positive iDANs. Here we systematically refine the culture conditions used for dopaminergic reprogramming of human stem cell-derived glial progenitor cells (GPCs) and adult dermal fibroblasts (DFs). Starting from a standard reprogramming medium containing base medium, growth factors and small molecules, we systematically compared three base media and removed individual small molecules to determine their specific contribution to reprogramming outcome. The GPCs and DFs were reprogrammed using established transcription factor-based strategies combined with REST inhibition. We found that the two starting cell types benefited from different culture conditions. For GPCs, B27/Neurobasal medium without small molecules yielded the highest neuronal and TH reprogramming efficiency, whereas DFs yielded the highest efficiencies in N2/DMEM/Neurobasal containing small molecules. Further characterization of the GPC-derived iDANs showed that these cells were functional, expressed key dopaminergic markers, and released dopamine upon stimulation. Together, these findings demonstrate that media composition is a key parameter influencing dopaminergic reprogramming efficiency across starting cell types in vitro. (© 2026. The Author(s).) |
| Competing Interests: | Declarations. Competing interests: MP is inventor of patents relating to reprogramming skin cells to DA neurons. The other authors declare that they have no competing interests. |
| References: | Cell Res. 2012 Feb;22(2):321-32. (PMID: 22105488) Cell Syst. 2025 Apr 16;16(4):101205. (PMID: 40086434) Stem Cell Reports. 2022 Oct 11;17(10):2203-2219. (PMID: 36150382) Nat Biotechnol. 2017 May;35(5):444-452. (PMID: 28398344) Trends Mol Med. 2019 Oct;25(10):897-914. (PMID: 31371156) Cell Mol Life Sci. 2017 Oct;74(19):3553-3575. (PMID: 28698932) Protein Cell. 2017 May;8(5):328-348. (PMID: 28213718) Curr Protoc. 2022 Sep;2(9):e555. (PMID: 36121202) Nature. 2010 Feb 25;463(7284):1035-41. (PMID: 20107439) Nature. 2011 Jul 13;476(7359):228-31. (PMID: 21753754) Nat Methods. 2020 Nov;17(11):1156-1166. (PMID: 33087905) Nat Protoc. 2015 Oct;10(10):1543-55. (PMID: 26379228) Cells. 2021 Dec 08;10(12):. (PMID: 34943958) Nat Protoc. 2017 Sep;12(9):1962-1979. (PMID: 28858290) Cell Stem Cell. 2015 Dec 3;17(6):719-734. (PMID: 26526726) Curr Opin Genet Dev. 2021 Oct;70:76-82. (PMID: 34175529) Cell Rep. 2012 Jun 28;1(6):703-14. (PMID: 22813745) Nat Methods. 2012 Jun;9(6):575-8. (PMID: 22484851) Nat Biotechnol. 2020 Dec;38(12):1421-1430. (PMID: 33273741) Commun Biol. 2022 Jun 10;5(1):578. (PMID: 35689020) Nat Neurosci. 2002 Apr;5(4):308-15. (PMID: 11896398) Nat Rev Neurol. 2015 Sep;11(9):492-503. (PMID: 26240036) Cell Stem Cell. 2015 Dec 3;17(6):735-747. (PMID: 26481520) Hum Gene Ther. 2004 Oct;15(10):934-44. (PMID: 15585109) Nature. 2010 Nov 11;468(7321):214-22. (PMID: 21068830) Neuron. 2024 Apr 3;112(7):1117-1132.e9. (PMID: 38266647) J Vis Exp. 2018 Feb 5;(132):. (PMID: 29443113) Nat Neurosci. 2018 Jul;21(7):932-940. (PMID: 29915193) Sci Rep. 2014 Sep 11;4:6330. (PMID: 25208484) Stem Cell Reports. 2020 Oct 13;15(4):869-882. (PMID: 32976765) FEBS Lett. 2019 Dec;593(23):3370-3380. (PMID: 31535361) Nature. 2017 Apr 13;544(7649):245-249. (PMID: 28379941) Sci Rep. 2016 Dec 05;6:38290. (PMID: 27917895) J Cell Biol. 2010 Apr 5;189(1):159-70. (PMID: 20351065) PLoS Biol. 2010 May 18;8(5):e1000373. (PMID: 20502524) Cells. 2020 Nov 10;9(11):. (PMID: 33182669) Cell Stem Cell. 2015 Dec 3;17(6):705-718. (PMID: 26456686) Proc Natl Acad Sci U S A. 2011 Jun 21;108(25):10343-8. (PMID: 21646515) Cell Syst. 2025 Apr 16;16(4):101206. (PMID: 40086435) STAR Protoc. 2025 Dec 19;6(4):104088. (PMID: 40946308) Nat Commun. 2013;4:2183. (PMID: 23873306) Nat Biotechnol. 1997 Sep;15(9):871-5. (PMID: 9306402) Science. 2024 Aug 2;385(6708):adl2992. (PMID: 39088624) Nature. 2011 Jul 03;476(7359):224-7. (PMID: 21725324) Cell Stem Cell. 2015 Aug 6;17(2):204-12. (PMID: 26253202) EMBO Mol Med. 2017 Aug;9(8):1117-1131. (PMID: 28646119) Sci Adv. 2025 Jun 6;11(23):eadq7855. (PMID: 40479059) Stem Cell Reports. 2017 Sep 12;9(3):742-751. (PMID: 28844658) Cell Stem Cell. 2012 Oct 5;11(4):471-6. (PMID: 23040476) Cell Rep Methods. 2024 Sep 16;4(9):100845. (PMID: 39236715) |
| Contributed Indexing: | Keywords: Culture conditions; Direct fibroblast to neuron reprogramming; Direct glia to neuron reprogramming; Induced dopaminergic neurons; Media composition |
| Substance Nomenclature: | 0 (Culture Media) EC 1.14.16.2 (Tyrosine 3-Monooxygenase) VTD58H1Z2X (Dopamine) |
| Entry Date(s): | Date Created: 20260817 Date Completed: 20260818 Latest Revision: 20260831 |
| Update Code: | 20260831 |
| PubMed Central ID: | PMC13483191 |
| DOI: | 10.1038/s41598-026-66620-z |
| PMID: | 42608437 |
| Βάση Δεδομένων: | MEDLINE |
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