Abstract
Oligodendrocytes are the main glial cell type in the central nervous system supporting the axonal part of neurons via myelin and lactate delivery. Both the conductive myelin formation and the energy support via lactate can be affected in diseases, such as multiple sclerosis and amyotrophic lateral sclerosis, respectively. Therefore, human disease modeling is needed to gain more mechanistic insights to drive drug discovery research. Here, patient-derived induced pluripotent stem cells (iPSCs) serve as a necessary tool providing an infinite cell source for patient-specific disease modeling, which allows investigation of oligodendrocyte involvement in human disease.
Small molecule-based differentiation protocols to generate oligodendrocytes from pluripotent stem cells can last more than 90 days. Here, we provide a transcription factor-based, fast and efficient protocol for generating O4+ oligodendrocytes in just 20–24 days. After a neural induction phase of 8–12 days, SOX10 is overexpressed either with the use of lentiviral vectors or via engineered iPSCs, which inducibly overexpress SOX10 after doxycycline addition. Using this last method, a pure O4+ cell population is achieved after keeping the SOX10-overexpressing neural stem cells in culture for an additional 10 days. Furthermore, these O4+ cells can be co-cultured with iPSC-derived cortical neurons in 384-well format, allowing pro-myelinating drug screens. In conclusion, we provide a fast and efficient oligodendrocyte differentiation protocol allowing both in vitro human disease modeling and a high-throughput co-culture system for drug discovery.
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References
Trapp BD, Nave K (2008) Multiple sclerosis: an immune or neurodegenerative disorder? Annu Rev Neurosci 31:247–269
Philips T, Rothstein JD (2017) Oligodendroglia: metabolic supporters of neurons. J Clin Invest 127(9):3271–3280
Lee Y, Morrison BM, Li Y, Lengacher S, Farah MH, Hoffman PN et al (2012) Oligodendroglia metabolically support axons and contribute to neurodegeneration. Nature 487(7408):443–448
Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K et al (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131(5):861–872
Chambers SM, Fasano CA, Papapetrou EP, Tomishima M, Sadelain M, Studer L (2009) Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol 27(3):275–280
Goldman SA, Kuypers NJ (2015) How to make an oligodendrocyte. Development 142(23):3983–3995
Gaspard N, Vanderhaeghen P (2010) Mechanisms of neural specification from embryonic stem cells. Curr Opin Neurobiol 20(1):37–43
Wang S, Bates J, Li X, Schanz S, Chandler-militello D, Levine C et al (2013) Clinical progress progenitor cells can myelinate and rescue a mouse model of congenital hypomyelination. Stem Cell 12(2):252–264
Livesey MR, Magnani D, Cleary EM, Vasistha NA, James OT, Selvaraj BT et al (2016) Maturation and electrophysiological properties of human pluripotent stem cell-derived oligodendrocytes. Stem Cells 34(4):1040–1053
Douvaras P, Wang J, Zimmer M, Hanchuk S, O’Bara MA, Sadiq S et al (2014) Efficient generation of myelinating oligodendrocytes from primary progressive multiple sclerosis patients by induced pluripotent stem cells. Stem Cell Reports 3(2):250–259
Ehrlich M, Mozafari S, Glatza M, Starost L, Velychko S, Hallmann A-L et al (2017) Rapid and efficient generation of oligodendrocytes from human induced pluripotent stem cells using transcription factors. Proc Natl Acad Sci 114(11):E2243–E2252
García-León JA, Kumar M, Boon R, Chau D, One J, Wolfs E et al (2018) SOX10 single transcription factor-based fast and efficient generation of oligodendrocytes from human pluripotent stem cells. Stem Cell Reports 10(2):655–672
Ordovás L, Boon R, Pistoni M, Chen Y, Wolfs E, Guo W et al (2015) Efficient recombinase-mediated cassette exchange in hPSCs to study the hepatocyte lineage reveals AAVS1 locus-mediated transgene inhibition. Stem Cell Reports 5(5):918–931
Marti E, Bumcrot DA, Takada R, Mcmahon AP (1995) Requirement of 19K form of Sonic hedgehog for induction of distinct ventral cell types in CNS explants. Nature 375(May):322–325
Shi Y, Kirwan P, Livesey FJ (2012) Directed differentiation of human pluripotent stem cells to cerebral cortex neurons and neural networks. Nat Protoc 7(10):1836–1846
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Neyrinck, K., García-León, J.A. (2021). Single Transcription Factor-Based Differentiation Allowing Fast and Efficient Oligodendrocyte Generation via SOX10 Overexpression. In: Ahlenius, H. (eds) Neural Reprogramming. Methods in Molecular Biology, vol 2352. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1601-7_11
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DOI: https://doi.org/10.1007/978-1-0716-1601-7_11
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1600-0
Online ISBN: 978-1-0716-1601-7
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