Abstract
Human brain organoids provide opportunities to produce three-dimensional (3D) brain-like tissues for biomedical research and translational drug discovery, toxicology, and tissue replacement. Here we describe a protocol for rapid and defined induction of brain organoids from human induced pluripotent stem cells (iPSCs), using commercially available culture and differentiation media and a cheap, easy to handle and clinically approved semisynthetic hydrogel. Importantly, the methodology is uncomplicated, well-defined, and reliable for reproducible and scalable organoid generation, and amendable to principles of current good laboratory practice (cGLP), with the potential for prospective adaptation to current good manufacturing practice (cGMP) toward clinical compliance.
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References
Lancaster MA, Knoblich JA (2014) Generation of cerebral organoids from human pluripotent stem cells. Nat Protoc 9:2329–2340
Lancaster MA, Renner M, Martin CA, Wenzel D, Bicknell LS, Hurles ME et al (2013) Cerebral organoids model human brain development and microcephaly. Nature 501:373–379
Li Y, Muffat J, Omer A, Bosch I, Lancaster MA, Sur M et al (2017) Induction of expansion and folding in human cerebral organoids. Cell Stem Cell 20:385–396 e3
Lindborg BA, Brekke JH, Vegoe AL, Ulrich CB, Haider KT, Subramaniam S et al (2016) Rapid induction of cerebral organoids from human induced pluripotent stem cells using a chemically defined hydrogel and defined cell culture medium. Stem Cells Transl Med 5:970–979
Crook JM, Ludwig TE (eds) (2017) Stem cell banking: concepts and protocols. methods in molecular biology, vol 1590. Springer, New York
Andrews PW, Baker D, Benvinisty N, Miranda B, Bruce K, Brustle O et al (2015) Points to consider in the development of seed stocks of pluripotent stem cells for clinical applications: international stem cell banking initiative (ISCBI). Regen Med 10:1–44
International Stem Cell Banking I (2009) Consensus guidance for banking and supply of human embryonic stem cell lines for research purposes. Stem Cell Rev 5:301–314
Yue K, Trujillo-de Santiago G, Alvarez MM, Tamayol A, Annabi N, Khademhosseini A (2015) Synthesis, properties, and biomedical applications of gelatin methacryloyl (GelMA) hydrogels. Biomaterials 73:254–271
Brehm JL, Ludwig TE (2017) Culture, adaptation, and expansion of pluripotent stem cells. In: Crook JM, Ludwig TE (eds) Stem cell banking: concepts and protocols. Methods in molecular biology, vol 1590. pp 139–150
Nichol JW, Koshy ST, Bae H, Hwang CM, Yamanlar S, Khademhosseini A (2010) Cell-laden microengineered gelatin methacrylate hydrogels. Biomaterials 31:5536–5544
O’Connell CD, Di Bella C, Thompson F, Augustine C, Beirne S, Cornock R et al (2016) Development of the Biopen: a handheld device for surgical printing of adipose stem cells at a chondral wound site. Biofabrication 8:015019
Acknowledgment
The authors wish to acknowledge funding from the Australian Research Council (ARC) Centre of Excellence Scheme (CE140100012).
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Tomaskovic-Crook, E., Crook, J.M. (2017). Clinically Amendable, Defined, and Rapid Induction of Human Brain Organoids from Induced Pluripotent Stem Cells. In: Turksen, K. (eds) Organoids. Methods in Molecular Biology, vol 1576. Humana, New York, NY. https://doi.org/10.1007/7651_2017_95
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DOI: https://doi.org/10.1007/7651_2017_95
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Publisher Name: Humana, New York, NY
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