Abstract
Neural crest cells (NCCs) are a multipotent and transient cell population that gives rise to many important tissues during human embryogenesis. Disturbances that occur during NCCs development may lead to numerous types of diseases and syndromes, which are called neurocristopathies. NCCs in vitro modeling enables the access to cellular, genetic, and biochemical information about the neural crest development and its derivatives. By using cells derived from patients with neurocristopathies it is possible to study the cellular and genetic mechanisms behind each disease in a specific and trustworthy manner, as well as to contribute to the development of prospective treatments. Here, we describe a protocol of 19 days, capable of efficiently generating NCCs from human induced pluripotent stem cells (hiPSCs). This differentiation process recapitulates the intermediate stage of neural plate border-like cells (NBCs), the epithelial to mesenchymal transition (EMT), and enables further generation of NCCs derivatives, such as Schwann cells, smooth muscle cells, melanocytes, peripheral neurons, adipocytes, osteoblasts, and chondrocytes.
Diogo Andrade Nani and Gabriella Shih Ping Hsia are co-first authors.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hall BK (2008) The neural crest and neural crest cells in vertebrate development and evolution, vol 11. Springer Science & Business Media, Berlin
Trainor P (2014) Neural crest cells evolution, development and disease. Elsevier, Amsterdam. ISBN: 978-0-12-401730-6
Luksch R, Castellani MR, Collini P, De Bernardi B, Conte M, Gambini C et al (2016) Neuroblastoma (peripheral neuroblastic tumours). Crit Rev Oncol Hematol 107:163–181
Nakamura T (1995) Genetic markers and animal models of neurocristopathy. Histol Histopathol 10(3):747–759
Lippmann ES, Estevez-Silva MC, Ashton RS (2014) Defined human pluripotent stem cell culture enables highly efficient neuroepithelium derivation without small molecule inhibitors. Stem Cells 32(4):1032–1042
Roellig D, Tan-Cabugao J, Esaian S, Bronner ME (2017) Dynamic transcriptional signature and cell fate analysis reveals plasticity of individual neural plate border cells. eLife 6:e21620
Maharana SK, Schlosser G (2018) A gene regulatory network underlying the formation of pre-placodal ectoderm in Xenopus laevis. BMC Biol 16(1):1–18
Kobayashi GS, Musso CM, de Paula Moreira D, Pontillo-Guimarães G, Hsia GSP, Caires-Júnior LC et al (2020) Recapitulation of neural crest specification and EMT via induction from neural plate border-like cells. Stem Cell Rep 15(3):776–788
Okita K, Matsumura Y, Sato Y, Okada A, Morizane A, Okamoto S et al (2011) A more efficient method to generate integration-free human iPS cells. Nat Methods 8(5):409–412
Riento K, Ridley AJ (2003) Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 4(6):446–456
Thermo Fisher Scientific. (2009). Handle with care: gentle dissociation product for sensitive cells [StemPro® Accutase® cell dissociation reagent]. Invitrogen. https://tools.thermofisher.com/content/sfs/brochures/F-081356_Accutase_Flyer_FHR.pdf. Accessed 14 May 2021
Kleinman HK, McGarvey ML, Liotta LA, Robey PG, Tryggvason K, Martin GR (1982) Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry 21(24):6188–6193
Kleinman HK, McGarvey ML, Hassell JR, Star VL, Cannon FB, Laurie GW, Martin GR (1986) Basement membrane complexes with biological activity. Biochemistry 25(2):312–318
Sigma-Aldrich (2009). Product information CryoStor CS2, CS5, and CS10 Cryopreservation Media. BioLife Solutions, Inc. https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Bulletin/1/c2874bul.pdf. Accessed 14 May 2021
Soldatov R, Kaucka M, Kastriti ME, Petersen J, Chontorotzea T, Englmaier L et al (2019) Spatiotemporal structure of cell fate decisions in murine neural crest. Science 364(6444)
Xue X, Sun Y, Resto-Irizarry AM, Yuan Y, Yong KMA, Zheng Y et al (2018) Mechanics-guided embryonic patterning of neuroectoderm tissue from human pluripotent stem cells. Nat Mater 17(7):633–641
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Nani, D.A., Hsia, G.S.P., Passos-Bueno, M.R., Kobayashi, G.S. (2021). Modeling Early Neural Crest Development via Induction from hiPSC-Derived Neural Plate Border-like Cells. In: Turksen, K. (eds) Induced Pluripotent Stem Cells and Human Disease. Methods in Molecular Biology, vol 2549. Humana, New York, NY. https://doi.org/10.1007/7651_2021_454
Download citation
DOI: https://doi.org/10.1007/7651_2021_454
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2584-2
Online ISBN: 978-1-0716-2585-9
eBook Packages: Springer Protocols