Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients

Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows in-depth assessment of transcriptional changes in immune cells of patients with COVID-19. However, collecting, processing, and analyzing samples from patients with COVID-19 pose many challenges because blood...

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Detalles Bibliográficos
Autores principales: Yao, Changfu, Bora, Stephanie A., Chen, Peter, Goodridge, Helen S., Gharib, Sina A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8114807/
https://www.ncbi.nlm.nih.gov/pubmed/34002169
http://dx.doi.org/10.1016/j.xpro.2021.100582
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author Yao, Changfu
Bora, Stephanie A.
Chen, Peter
Goodridge, Helen S.
Gharib, Sina A.
author_facet Yao, Changfu
Bora, Stephanie A.
Chen, Peter
Goodridge, Helen S.
Gharib, Sina A.
author_sort Yao, Changfu
collection PubMed
description Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows in-depth assessment of transcriptional changes in immune cells of patients with COVID-19. However, collecting, processing, and analyzing samples from patients with COVID-19 pose many challenges because blood samples may contain infectious virus, identification of immune cell subtypes can be difficult, and biological interpretation of analytical results is complex. To address these issues, we describe a protocol for sample processing, sorting, methanol fixation, and scRNA-seq analysis of PBMCs from frozen buffy coat samples from patients with COVID-19. For complete details on the use and execution of this protocol, please refer to (Yao et al., 2021).
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spelling pubmed-81148072021-05-13 Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients Yao, Changfu Bora, Stephanie A. Chen, Peter Goodridge, Helen S. Gharib, Sina A. STAR Protoc Protocol Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows in-depth assessment of transcriptional changes in immune cells of patients with COVID-19. However, collecting, processing, and analyzing samples from patients with COVID-19 pose many challenges because blood samples may contain infectious virus, identification of immune cell subtypes can be difficult, and biological interpretation of analytical results is complex. To address these issues, we describe a protocol for sample processing, sorting, methanol fixation, and scRNA-seq analysis of PBMCs from frozen buffy coat samples from patients with COVID-19. For complete details on the use and execution of this protocol, please refer to (Yao et al., 2021). Elsevier 2021-05-12 /pmc/articles/PMC8114807/ /pubmed/34002169 http://dx.doi.org/10.1016/j.xpro.2021.100582 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Yao, Changfu
Bora, Stephanie A.
Chen, Peter
Goodridge, Helen S.
Gharib, Sina A.
Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients
title Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients
title_full Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients
title_fullStr Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients
title_full_unstemmed Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients
title_short Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients
title_sort sample processing and single cell rna-sequencing of peripheral blood immune cells from covid-19 patients
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8114807/
https://www.ncbi.nlm.nih.gov/pubmed/34002169
http://dx.doi.org/10.1016/j.xpro.2021.100582
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