Organoids pp 13-22 | Cite as

Clinically Amendable, Defined, and Rapid Induction of Human Brain Organoids from Induced Pluripotent Stem Cells

  • Eva Tomaskovic-CrookEmail author
  • Jeremy M. CrookEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1576)


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.


Brain organoid Clinical compliance Defined GelMA Human induced pluripotent stem cells Hydrogel 



The authors wish to acknowledge funding from the Australian Research Council (ARC) Centre of Excellence Scheme (CE140100012).


  1. 1.
    Lancaster MA, Knoblich JA (2014) Generation of cerebral organoids from human pluripotent stem cells. Nat Protoc 9:2329–2340CrossRefGoogle Scholar
  2. 2.
    Lancaster MA, Renner M, Martin CA, Wenzel D, Bicknell LS, Hurles ME et al (2013) Cerebral organoids model human brain development and microcephaly. Nature 501:373–379CrossRefGoogle Scholar
  3. 3.
    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 e3CrossRefGoogle Scholar
  4. 4.
    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 5:970–979CrossRefGoogle Scholar
  5. 5.
    Crook JM, Ludwig TE (eds) (2017) Stem cell banking: concepts and protocols. methods in molecular biology, vol 1590. Springer, New YorkGoogle Scholar
  6. 6.
    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 10:1–44CrossRefGoogle Scholar
  7. 7.
    International Stem Cell Banking I (2009) Consensus guidance for banking and supply of human embryonic stem cell lines for research purposes. Stem Cell Rev 5:301–314CrossRefGoogle Scholar
  8. 8.
    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 73:254–271CrossRefGoogle Scholar
  9. 9.
    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 139–150Google Scholar
  10. 10.
    Nichol JW, Koshy ST, Bae H, Hwang CM, Yamanlar S, Khademhosseini A (2010) Cell-laden microengineered gelatin methacrylate hydrogels. Biomaterials 31:5536–5544CrossRefGoogle Scholar
  11. 11.
    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 8:015019CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.ARC Centre of Excellence for Electromaterials Science, Intelligent Polymer Research Institute, AIIM Facility, Innovation CampusUniversity of WollongongWollongongAustralia
  2. 2.Illawarra Health and Medical Research InstituteUniversity of WollongongWollongongAustralia
  3. 3.Department of Surgery, St Vincent’s HospitalThe University of MelbourneFitzroyAustralia

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