Abstract
Human-induced pluripotent stem cells (hiPSCs) are useful tools to examine human neuronal maturation processes. In this chapter, we describe the maturation of human neuronal precursor cells derived from hiPSCs by cellular communication network family member 2, also known as connective tissue growth factor. We describe the (1) preparation of feeder cells for undifferentiated culture of hiPSCs, (2) undifferentiated culture of hiPSCs, (3) induction of neuronal precursor cells from hiPSCs, (4) maturation of neuronal precursor cells from hiPSCs, (5) immunofluorescent staining of neuronal cells from hiPSCs, and (6) immunofluorescence analysis.
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
Takigawa M (2013) CCN2: a master regulator of the genesis of bone and cartilage. J Cell Commun Signal 7(3):191–201. https://doi.org/10.1007/s12079-013-0204-8
Takigawa M (2018) An early history of CCN2/CTGF research: the road to CCN2 via hcs24, ctgf, ecogenin, and regenerin. J Cell Commun Signal 12(1):253–264. https://doi.org/10.1007/s12079-017-0414-6
Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4):663–676. https://doi.org/10.1016/j.cell.2006.07.024
Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131(5):861–872. https://doi.org/10.1016/j.cell.2007.11.019
Shoji M, Nishioka M, Minato H, Harada K, Kubo M, Fukuyama Y, Kuzuhara T (2016) Neurotrophic activity of jiadifenolide on neuronal precursor cells derived from human induced pluripotent stem cells. Biochem Biophys Res Commun 470(4):798–803. https://doi.org/10.1016/j.bbrc.2016.01.092
Shoji M, Ueda M, Nishioka M, Minato H, Seki M, Harada K, Kubo M, Fukuyama Y, Suzuki Y, Aoyama E, Takigawa M, Kuzuhara T (2019) Jiadifenolide induces the expression of cellular communication network factor (CCN) genes, and CCN2 exhibits neurotrophic activity in neuronal precursor cells derived from human induced pluripotent stem cells. Biochem Biophys Res Commun 519(2):309–315. https://doi.org/10.1016/j.bbrc.2019.09.003
Eiraku M, Sasai Y (2012) Mouse embryonic stem cell culture for generation of three-dimensional retinal and cortical tissues. Nat Protoc 7(1):69–79. https://doi.org/10.1038/nprot.2011.429
Kondo T, Asai M, Tsukita K, Kutoku Y, Ohsawa Y, Sunada Y, Imamura K, Egawa N, Yahata N, Okita K, Takahashi K, Asaka I, Aoi T, Watanabe A, Watanabe K, Kadoya C, Nakano R, Watanabe D, Maruyama K, Hori O, Hibino S, Choshi T, Nakahata T, Hioki H, Kaneko T, Naitoh M, Yoshikawa K, Yamawaki S, Suzuki S, Hata R, Ueno S, Seki T, Kobayashi K, Toda T, Murakami K, Irie K, Klein WL, Mori H, Asada T, Takahashi R, Iwata N, Yamanaka S, Inoue H (2013) Modeling Alzheimer's disease with iPSCs reveals stress phenotypes associated with intracellular Abeta and differential drug responsiveness. Cell Stem Cell 12(4):487–496. https://doi.org/10.1016/j.stem.2013.01.009
Pool M, Thiemann J, Bar-Or A, Fournier AE (2008) NeuriteTracer: a novel ImageJ plugin for automated quantification of neurite outgrowth. J Neurosci Methods 168(1):134–139. https://doi.org/10.1016/j.jneumeth.2007.08.029
Watanabe K, Ueno M, Kamiya D, Nishiyama A, Matsumura M, Wataya T, Takahashi JB, Nishikawa S, Nishikawa S, Muguruma K, Sasai Y (2007) A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol 25(6):681–686. https://doi.org/10.1038/nbt1310
Acknowledgments
This work was supported by the Japan Society for the Promotion of Science (JSPS), Grants-in-Aid for Scientific Research (C) 18K06645 (to T.K.), Grants-in-Aid for Young Scientists (B) 26860083 (to M.S.), and the Tokushima Bunri University Grant for Educational Reform and the Collaborative Research Grants TBU2015-2-2 (to M.S.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Shoji, M., Kuzuhara, T. (2023). Imaging Analysis of Neurotrophic Effects by CCN2 Protein in Neuronal Precursor Cells Derived from Human-Induced Pluripotent Stem Cells. In: Takigawa, M. (eds) CCN Proteins. Methods in Molecular Biology, vol 2582. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2744-0_18
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2744-0_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2743-3
Online ISBN: 978-1-0716-2744-0
eBook Packages: Springer Protocols