Infecting human hematopoietic stem and progenitor cells with SARS-CoV-2

• Published in: STAR protocols Vol. 2; no. 4; p. 100903
• Main Authors: Huerga Encabo, Hector, Ulferts, Rachel, Sharma, Aneesh, Beale, Rupert, Bonnet, Dominique
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17-12-2021; Elsevier
• Subjects: Bone Marrow - virology; Cell Biology; Cell isolation; Cell-based Assays; Colony-Forming Units Assay - methods; COVID-19 - pathology; COVID-19 - virology; COVID-19 Nucleic Acid Testing - methods; Fetal Blood - virology; Hematopoietic Stem Cells - pathology; Hematopoietic Stem Cells - virology; Humans; Immunology; Microbiology; Microscopy; Protocol; SARS-CoV-2 - isolation & purification; Stem Cells; Stem Cells; Immunology; Microbiology; Microscopy; Cell-based Assays; Cell isolation; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100903

Determining how hematopoietic stem and progenitor cells (HSPCs) can be infected by viruses is necessary to understand and predict how the immune system will drive the host response. We present here a protocol to analyze the capacity of SARS-CoV-2 to infect different subsets of human HSPCs, inlcuding procedures for SARS-CoV-2 production and titration, isolation of human HSPCs from different sources (bone marrow, umbilical cord, or peripheral blood), and quantification of SARS-Cov-2 infection capacity by RT-qPCR and colony forming unit assay.
For complete details on the use and execution of this protocol, please refer to Huerga Encabo et al. (2021)
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•Production and titration of SARS-CoV-2•Isolation of human hematopoietic stem and progenitor cells from different sources•Analysis of the infection capacity of SARS-CoV-2 in HSPCs•Despite their intrinsic antiviral protection HSPCs can be infected by SARS-CoV-2
Determining how hematopoietic stem and progenitor cells (HSPCs) can be infected by viruses is necessary to understand and predict how the immune system will drive the host response. We present here a protocol to analyze the capacity of SARS-CoV-2 to infect different subsets of human HSPCs, inlcuding procedures for SARS-CoV-2 production and titration, isolation of human HSPCs from different sources (bone marrow, umbilical cord or peripheral blood), and quantification of SARS-Cov-2 infection capacity by RT-PCR and colony forming unit assay.