Brain-Region-Specific Organoids Using Mini-bioreactors for Modeling ZIKV Exposure.
Cerebral organoids, three-dimensional cultures that model organogenesis, provide a new platform to investigate human brain development. High cost, variability, and tissue heterogeneity limit their broad applications. Here, we developed a miniaturized spinning bioreactor (SpinΩ) to generate forebrain-specific organoids from human iPSCs. These organoids recapitulate key features of human cortical development, including progenitor zone organization, neurogenesis, gene expression, and, notably, a distinct human-specific outer radial glia cell layer. We also developed protocols for midbrain and hypothalamic organoids. Finally, we employed the forebrain organoid platform to model Zika virus (ZIKV) exposure. Quantitative analyses revealed preferential, productive infection of neural progenitors with either African or Asian ZIKV strains. ZIKV infection leads to increased cell death and reduced proliferation, resulting in decreased neuronal cell-layer volume resembling microcephaly. Together, our brain-region-specific organoids and SpinΩ provide an accessible and versatile platform for modeling human brain development and disease and for compound testing, including potential ZIKV antiviral drugs.
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Biomedical Engineering Graduate Program, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Xuyu
X
Qian
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Graduate Program in Cellular and Molecular Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Ha Nam
HN
Nguyen
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Riverhill High School, Clarksville, MD 21029, USA.
Mingxi M
MM
Song
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Byram Hills High School, Armonk, NY 10504, USA.
Christopher
C
Hadiono
Department of Biological Science, Florida State University, Tallahassee, FL 32306, USA.
Sarah C
SC
Ogden
Department of Biological Science, Florida State University, Tallahassee, FL 32306, USA.
Christy
C
Hammack
Department of Human Genetics, School of Medicine, Emory University, Atlanta, GA 30322, USA.
Bing
B
Yao
Lieber Institute for Brain Development, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Gregory R
GR
Hamersky
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Fadi
F
Jacob
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Chun
C
Zhong
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Ki-Jun
KJ
Yoon
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Clear Lake High School, Harris, TX 77058, USA.
William
W
Jeang
Department of Human Genetics, School of Medicine, Emory University, Atlanta, GA 30322, USA.
Li
L
Lin
Department of Human Genetics, School of Medicine, Emory University, Atlanta, GA 30322, USA.
Yujing
Y
Li
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Jai
J
Thakor
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Daniel A
DA
Berg
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Ce
C
Zhang
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Eunchai
E
Kang
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Michael
M
Chickering
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
David
D
Nauen
Division of Pathology, Children's National Medical Center, George Washington University, Washington, DC 20010, USA; Pathology and Pediatrics, George Washington University, Washington, DC 20010, USA.
Cheng-Ying
CY
Ho
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Zhexing
Z
Wen
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Kimberly M
KM
Christian
Department of Biochemistry and Molecular Biology, Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX 77555, USA.
Pei-Yong
PY
Shi
Lieber Institute for Brain Development, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Brady J
BJ
Maher
Department of Biostatistics and Bioinformatics, Rollins School of Public Health, Emory University, Atlanta, GA 30322, USA.
Hao
H
Wu
Department of Human Genetics, School of Medicine, Emory University, Atlanta, GA 30322, USA.
Peng
P
Jin
Department of Biological Science, Florida State University, Tallahassee, FL 32306, USA.
Hengli
H
Tang
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Graduate Program in Cellular and Molecular Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Solomon Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: shongju1@jhmi.edu.
Hongjun
H
Song
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Graduate Program in Cellular and Molecular Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Solomon Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: gming1@jhmi.edu.
Guo-Li
GL
Ming
D019149:Bioreactors; D001921:Brain / Q000166:cytology*; D018929:Cell Culture Techniques / Q000191:economics; D004622:Embryo, Mammalian; D047108:Embryonic Development; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D008954:Models, Biological; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology; D009940:Organoids / Q000821:virology; D000071244:Zika Virus / Q000502:physiology*; D000071243:Zika Virus Infection / Q000503:physiopathology / Q000821:virology