Sliced Human Cortical Organoids for Modeling Distinct Cortical Layer Formation

Xuyu Qian; Yijing Su; Christopher D Adam; Andre U Deutschmann; Sarshan R Pather; Ethan M Goldberg; Kenong Su; Shiying Li; Lu Lu; Fadi Jacob; Phuong T T Nguyen; Sooyoung Huh; Ahmet Hoke; Sarah E Swinford-Jackson; Zhexing Wen; Xiaosong Gu; R Christopher Pierce; Hao Wu; Lisa A Briand; H Isaac Chen; John A Wolf; Hongjun Song; Guo-Li Ming

doi:10.1016/j.stem.2020.02.002

Sliced Human Cortical Organoids for Modeling Distinct Cortical Layer Formation

Cell Stem Cell. 2020 May 7;26(5):766-781.e9. doi: 10.1016/j.stem.2020.02.002. Epub 2020 Mar 5.

Authors

Xuyu Qian¹, Yijing Su², Christopher D Adam³, Andre U Deutschmann⁴, Sarshan R Pather⁵, Ethan M Goldberg⁶, Kenong Su⁷, Shiying Li⁸, Lu Lu², Fadi Jacob⁹, Phuong T T Nguyen³, Sooyoung Huh², Ahmet Hoke¹⁰, Sarah E Swinford-Jackson¹¹, Zhexing Wen¹², Xiaosong Gu¹³, R Christopher Pierce¹⁴, Hao Wu¹⁵, Lisa A Briand⁴, H Isaac Chen¹⁶, John A Wolf¹⁷, Hongjun Song¹⁸, Guo-Li Ming¹⁹

Affiliations

¹ Department of Neuroscience and Mahoney Institute for Neurosciences, University of Pennsylvania, Philadelphia, PA 19104, USA; Biomedical Engineering Graduate Program, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
² Department of Neuroscience and Mahoney Institute for Neurosciences, University of Pennsylvania, Philadelphia, PA 19104, USA.
³ Neuroscience Graduate Program, University of Pennsylvania, Philadelphia, PA 19104, USA.
⁴ Department of Psychology, Temple University, Philadelphia, PA 19122, USA.
⁵ Cell and Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, PA 19104, USA.
⁶ Department of Neuroscience and Mahoney Institute for Neurosciences, University of Pennsylvania, Philadelphia, PA 19104, USA; Division of Neurology, Department of Pediatrics, The Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
⁷ Department of Computer Science, Emory University College of Arts and Sciences, Atlanta, GA 30322, USA.
⁸ Department of Neuroscience and Mahoney Institute for Neurosciences, University of Pennsylvania, Philadelphia, PA 19104, USA; Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu 226001, China.
⁹ Department of Neuroscience and Mahoney Institute for Neurosciences, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
¹⁰ Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
¹¹ Department of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA.
¹² Department of Psychiatry and Behavioral Science, Emory University School of Medicine, Atlanta, GA 30322, USA; Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
¹³ Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu 226001, China.
¹⁴ Neuroscience Graduate Program, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA.
¹⁵ Department of Biostatistics and Bioinformatics, Rollins School of Public Health, Emory University, Atlanta, GA 30322, USA.
¹⁶ Neuroscience Graduate Program, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Neurosurgery, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Corporal Michael J. Crescenz Veterans Affairs Medical Center, Philadelphia, PA 19104, USA.
¹⁷ Neuroscience Graduate Program, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Biostatistics and Bioinformatics, Rollins School of Public Health, Emory University, Atlanta, GA 30322, USA; Corporal Michael J. Crescenz Veterans Affairs Medical Center, Philadelphia, PA 19104, USA.
¹⁸ Department of Neuroscience and Mahoney Institute for Neurosciences, University of Pennsylvania, Philadelphia, PA 19104, USA; Neuroscience Graduate Program, University of Pennsylvania, Philadelphia, PA 19104, USA; Cell and Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Cell and Developmental Biology, University of Pennsylvania, Philadelphia, PA 19104, USA; The Epigenetics Institute, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
¹⁹ Department of Neuroscience and Mahoney Institute for Neurosciences, University of Pennsylvania, Philadelphia, PA 19104, USA; Neuroscience Graduate Program, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Cell and Developmental Biology, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: gming@pennmedicine.upen.edu.

Abstract

Human brain organoids provide unique platforms for modeling development and diseases by recapitulating the architecture of the embryonic brain. However, current organoid methods are limited by interior hypoxia and cell death due to insufficient surface diffusion, preventing generation of architecture resembling late developmental stages. Here, we report the sliced neocortical organoid (SNO) system, which bypasses the diffusion limit to prevent cell death over long-term cultures. This method leads to sustained neurogenesis and formation of an expanded cortical plate that establishes distinct upper and deep cortical layers for neurons and astrocytes, resembling the third trimester embryonic human neocortex. Using the SNO system, we further identify a critical role of WNT/β-catenin signaling in regulating human cortical neuron subtype fate specification, which is disrupted by a psychiatric-disorder-associated genetic mutation in patient induced pluripotent stem cell (iPSC)-derived SNOs. These results demonstrate the utility of SNOs for investigating previously inaccessible human-specific, late-stage cortical development and disease-relevant mechanisms.

Keywords: Brain organoid; DISC1; WNT; cerebral cortex; forebrain organoid; human iPSC; lamination; neurodevelopment; neuron fate specification; schizophrenia.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Humans
Induced Pluripotent Stem Cells*
Neocortex*
Neurogenesis
Neurons
Organoids

Abstract

Publication types

MeSH terms

Grants and funding