Phenotypic Profiling of PBMCs and iPSC-Derived Cells

Phenotypic analysis of PBMCs and human iPSC-derived cells was performed as described.22 24 27 (link) Briefly, single cell suspensions of PBMCs and human iPSC-derived cells were stained with a master mix of antibodies (Abs) for surface stains after Fc block (human Fc block; BD Biosciences) for 20 min at room temperature. Antibodies used in this study are listed in online supplemental table S2. Live/Dead Fixable Aqua Dead Cell Stain Kit (ThermoFisher Scientific) and LIVE/DEAD Fixable Near-IR Dead Cell Stain kit (ThermoFisher Scientific) were used to exclude dead cells from the analysis. Samples were analyzed using LSRII or LSRFortessa (BD Biosciences) with FlowJo software (TreeStar).

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Makino K., Long M.D., Kajihara R., Matsueda S., Oba T., Kanehira K., Liu S, & Ito F. (2022). Generation of cDC-like cells from human induced pluripotent stem cells via Notch signaling. Journal for Immunotherapy of Cancer, 10(1), e003827.

Publication 2022

human Fc block Live/Dead Fixable Aqua Dead Cell Stain Kit LIVE/DEAD Fixable Near-IR Dead Cell Stain kit LSRFortessa

Antibodies Cells Human Ipsc Phenotypic Stain Stains

Corresponding Organization : University of Southern California

Other organizations : Akita University, Shinshu University

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Variable analysis

independent variables

Phenotypic analysis of PBMCs and human iPSC-derived cells

dependent variables

Measured outcomes from phenotypic analysis

control variables

Fc block (human Fc block; BD Biosciences) for 20 min at room temperature
Live/Dead Fixable Aqua Dead Cell Stain Kit (ThermoFisher Scientific) and LIVE/DEAD Fixable Near-IR Dead Cell Stain kit (ThermoFisher Scientific) to exclude dead cells from the analysis

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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