All samples were analyzed by flow cytometry using a FACSAriaTM III (BD biosciences, San Jose, CA, USA). CD45 (Clone 104) (Biolegend, San Diego, CA, USA), CD45.1 (Clone A20) (BD Biosciences), CD45.2 (Clone 104) (Biolegend), EPCR (CD201) (Clone eBio1560, eBioscience, San Diego, CA, USA), CD150 (Clone mShad150, eBioscience), and CD48 (Clone HM48–1, Biolegend) antibodies were used to enumerate CD45+EPCR(CD201)+CD48−CD150+ (E-SLAM) cells.[22 (link),23 (link)] MKs were sorted on the basis of CD41 expression (Clone MWReg30, Biolegend) and cell size.[15 (link)] CD45 (Clone 104) (Biolegend) and CD31 (Clone 390, BD biosciences) antibodies were used to isolate CD45−CD31+ marrow ECs.[24 (link)]
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Multiparametric flow cytometry for hematopoietic stem cells
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Corresponding Organization : Northport VA Medical Center
Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College
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Variable analysis
independent variables
- Flow cytometry using a FACSAriaTM III (BD biosciences, San Jose, CA, USA)
- CD45 (Clone 104) (Biolegend, San Diego, CA, USA), CD45.1 (Clone A20) (BD Biosciences), CD45.2 (Clone 104) (Biolegend), EPCR (CD201) (Clone eBio1560, eBioscience, San Diego, CA, USA), CD150 (Clone mShad150, eBioscience), and CD48 (Clone HM48–1, Biolegend) antibodies
- CD41 expression (Clone MWReg30, Biolegend) and cell size for sorting MKs
- CD45 (Clone 104) (Biolegend) and CD31 (Clone 390, BD biosciences) antibodies for isolating CD45−CD31+ marrow ECs
- Enumeration of CD45+EPCR(CD201)+CD48−CD150+ (E-SLAM) cells
- Sorting of MKs
- Isolation of CD45−CD31+ marrow ECs
- Not explicitly mentioned
- No positive or negative controls were explicitly mentioned in the provided information.
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