FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures. - National Genomics Data Center (CNCB-NGDC)

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FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures.

Sylvie Janssens, Michael Schotsaert, Lara Manganaro, Marion Dejosez, Viviana Simon, Adolfo García-Sastre, Thomas P Zwaka

Author Information

Sylvie Janssens: Huffington Center for Cell-Based Research, Black Family Stem Cell Institute, Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY.
Michael Schotsaert: Department of Microbiology, Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY.
Lara Manganaro: Department of Microbiology, Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY.
Marion Dejosez: Huffington Center for Cell-Based Research, Black Family Stem Cell Institute, Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY.
Viviana Simon: Department of Microbiology, Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY.
Adolfo García-Sastre: Department of Microbiology, Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY.
Thomas P Zwaka: Huffington Center for Cell-Based Research, Black Family Stem Cell Institute, Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY.

PMID: 30358129 DOI: 10.1002/cpsc.65

Organoids-or pluripotent stem cell-derived in vitro-grown simplified mini organs-have become a tremendously important model to study human organ development and disease. To restrict the noise inherent to the heterogeneous cell mixtures derived from organoid cultures, we developed a new technique of fluorescence-assisted cell sorting (FACS) of virus-infected cerebral organoid cultures. This method still includes the advantage of growing cells in a more natural environment than traditional cell culture, but now renders samples suitable for downstream cell type-specific multi-omics analyses. The protocol starts from stem cell-derived mature brain organoids and includes steps for: preparing the culture for viral infection, production of the viral stocks, FACS sample preparation, and gating and sorting implementation. The protocol has been developed for Zika virus infection, but can be extrapolated to other viruses or fluorescent marker expression as illustrated in an alternate protocol using a single-cycle lentivirus expressing a fluorescent reporter protein. © 2018 by John Wiley & Sons, Inc.

FACS HIV Zika cerebral organoids human ESC lentiviral vector virus

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R21 AI125236/NIAID NIH HHS
R21 AI129486/NIAID NIH HHS
U19 AI118610/NIAID NIH HHS

Brain

Cell Culture Techniques

Cell Separation

Cells, Cultured

Flow Cytometry

Human Embryonic Stem Cells

Humans

Neurons

Organoids

Zika Virus

Zika Virus Infection

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