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FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures
- Source :
- Current protocols in stem cell biology. 48(1)
- Publication Year :
- 2018
-
Abstract
- 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.
- Subjects :
- 0301 basic medicine
viruses
Cell
Human Embryonic Stem Cells
Cell Culture Techniques
Cell Separation
Biology
Virus
Article
Viral vector
03 medical and health sciences
0302 clinical medicine
medicine
Organoid
Humans
Cells, Cultured
Neurons
Zika Virus Infection
Brain
Cell Biology
General Medicine
Zika Virus
Cell sorting
Flow Cytometry
Embryonic stem cell
Cell biology
Organoids
030104 developmental biology
medicine.anatomical_structure
Cell culture
030217 neurology & neurosurgery
Developmental Biology
Cerebral organoid
Subjects
Details
- ISSN :
- 19388969
- Volume :
- 48
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Current protocols in stem cell biology
- Accession number :
- edsair.doi.dedup.....e72124f33d816b36d686b6fef6baa3ac