Generation of human brain region-specific organoids using a miniaturized spinning bioreactor

Nature Protocols
Xuyu QianGuo-Li Ming

Abstract

Human brain organoids, 3D self-assembled neural tissues derived from pluripotent stem cells, are important tools for studying human brain development and related disorders. Suspension cultures maintained by spinning bioreactors allow for the growth of large organoids despite the lack of vasculature, but commercially available spinning bioreactors are bulky in size and have low throughput. Here, we describe the procedures for building the miniaturized multiwell spinning bioreactor SpinΩ from 3D-printed parts and commercially available hardware. We also describe how to use SpinΩ to generate forebrain, midbrain and hypothalamus organoids from human induced pluripotent stem cells (hiPSCs). These organoids recapitulate key dynamic features of the developing human brain at the molecular, cellular and structural levels. The reduction in culture volume, increase in throughput and reproducibility achieved using our bioreactor and region-specific differentiation protocols enable quantitative modeling of brain disorders and compound testing. This protocol takes 14-84 d to complete (depending on the type of brain region-specific organoids and desired developmental stages), and organoids can be further maintained over 200 d. Competence with...Continue Reading

References

Sep 14, 2010·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Mardi S Byerly, Seth Blackshaw
Nov 12, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Seth BlackshawTomomi Shimogori
Feb 4, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Teruko DanjoYoshiki Sasai
Jul 7, 2011·Cell·Jan H LuiArnold R Kriegstein
Sep 3, 2013·Nature·Madeline A LancasterJuergen A Knoblich
Nov 28, 2013·Proceedings of the National Academy of Sciences of the United States of America·Taisuke KadoshimaYoshiki Sasai
Jul 19, 2014·Science·Madeline A Lancaster, Juergen A Knoblich
Dec 27, 2015·Current Opinion in Neurobiology·Zhexing WenGuo-li Ming
Jan 11, 2016·Cell Stem Cell·Xiaolei YinOren Levy
Mar 16, 2017·Development·Xuyu QianGuo-Li Ming
Apr 25, 2017·Nature Cell Biology·Ya-Wen ChenHans-Willem Snoeck
Apr 27, 2017·Nature·Fikri BireySergiu P Paşca
May 16, 2017·Nature Methods·Joshua A BagleyJuergen A Knoblich
Jun 1, 2017·Nature Biotechnology·Madeline A LancasterJuergen A Knoblich

❮ Previous
Next ❯

Citations

Aug 10, 2018·Developmental Dynamics : an Official Publication of the American Association of Anatomists·H Isaac ChenGuo-Li Ming
Oct 17, 2018·The Journal of General Virology·Lindomar José PenaAntonio Carlos de Freitas
Nov 27, 2018·Molecular Biology of the Cell·Emily S Wilson, Karen Newell-Litwa
Jan 4, 2019·Advanced Healthcare Materials·Wujin SunAli Khademhosseini
May 7, 2019·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Davis M Aasen, M Natalia Vergara
May 10, 2019·Wiley Interdisciplinary Reviews. Developmental Biology·Christina Kyrousi, Silvia Cappello
Jun 6, 2019·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Charles B WrightPaul A Sieving
Jul 17, 2019·PLoS Computational Biology·Aleksandra KarolakKatarzyna A Rejniak
Jul 3, 2019·Molecular Neurobiology·Mirza Ali Mofazzal JahromiMichael R Hamblin
Aug 25, 2019·Molecular Psychiatry·Karl GrenierPhedias Diamandis
Sep 13, 2019·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Jordan G SchnollKimberly M Christian
Feb 28, 2020·Molecular Biology of the Cell·Gabriella L RobertsonVivian Gama
May 19, 2020·Neurosurgery·Rachel BlueH Isaac Chen
Mar 15, 2020·Biology of the Cell·Véronique Marthiens, Renata Basto
Apr 7, 2020·Journal of Molecular Cell Biology·Juan HeDong Gao
Jul 30, 2020·The Neuroscientist : a Review Journal Bringing Neurobiology, Neurology and Psychiatry·Anuradha YadavRajnish Kumar Chaturvedi
Jun 26, 2018·Frontiers in Synaptic Neuroscience·Hansen Wang
Jul 23, 2019·Tissue Engineering. Part C, Methods·Michael A PhelanPeter I Lelkes
Jul 6, 2019·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Jay Gopalakrishnan
Dec 10, 2019·Oxidative Medicine and Cellular Longevity·Marcelina KierońMichalina Wężyk
Mar 3, 2020·Tissue Engineering and Regenerative Medicine·Marina V Pryzhkova, Philip W Jordan
Feb 6, 2020·Frontiers in Cellular Neuroscience·Katrin SimmnacherBeate Winner
May 7, 2020·Frontiers in Cell and Developmental Biology·Roberta Azzarelli
Jul 15, 2020·Molecular Autism·Wai Kit ChanJohn O Mason
Sep 22, 2020·Current Protocols in Stem Cell Biology·Antje Appelt-MenzelOle Pless
Jun 24, 2018·Bioengineering·Cláudia C MirandaJoaquim M S Cabral
Jan 24, 2019·Bioengineering·Eyal Karzbrun, Orly Reiner
Jul 12, 2019·Frontiers in Neuroscience·Evelyn ChukwurahSofia B Lizarraga
Apr 25, 2020·Tissue Engineering and Regenerative Medicine·Hye-Jin JeongTae-Eun Park
Dec 12, 2019·Frontiers in Synaptic Neuroscience·Jorge Ojeda, Ariel Ávila
Apr 15, 2020·Annals of Biomedical Engineering·Katya D'CostaMilica Radisic
Jun 13, 2020·Frontiers in Cell and Developmental Biology·Victor J T LinYu-Chieh Wang
Jun 1, 2019·Stem Cells International·Teresa P SilvaTiago G Fernandes
Apr 18, 2019·Development·Xuyu QianGuo-Li Ming
Dec 19, 2019·Frontiers in Bioengineering and Biotechnology·Jing SunYangzi Jiang
Aug 28, 2020·Frontiers in Cellular Neuroscience·Cristina Salado-ManzanoJosep M Canals
Apr 8, 2020·International Journal of Molecular Sciences·Shyh Jenn ChiaYin-Xia Chao
Aug 9, 2019·Molecular Psychiatry·Jay PenneyLi-Huei Tsai
Aug 6, 2020·Bio-design and Manufacturing·Jai ThakorAli Khademhosseini
Jan 1, 2019·Advanced Biosystems·Roza Vaez GhaemiVikramaditya G Yadav
Jun 3, 2020·Cell Death and Differentiation·Jaydeep Sidhaye, Jürgen A Knoblich
Oct 22, 2020·The International Journal of Neuroscience·Na JiaLina Sun

❮ Previous
Next ❯

Related Concepts

Related Feeds

3D Cellular Models of Brain and Neurodegeneration

Brain organoids are three-dimensional in vitro cellular models of the brain that can recapitulate many processes such as the neurodevelopment. In addition, these organoids can be combined with other cell types, such as neurons and astrocytes to study their interactions in assembloids. Disease processes can also be modeled by induced pluripotent stem cell-derived organoids and assembloids from patients with neurodegenerative disorders. Discover the latest research on the models here.

Bioreactors (ASM)

Bioreactors are important devices or systems that utilize living cells or enzymes for a chemical process. These devices carefully control and monitor the environment factors such as pH, nutrients level, and temperature, which can affect the yield in a given process. Discover the latest research here.

Brain Organoids in Disease Modeling

Brain organoids are three-dimensional cell culture models derived from human pluripotent stem cells. Since they resemble the embryonic brain, they can be used to help study brain biology, early brain development, and brain diseases. Discover the latest research on brain organoids in disease modeling here.