MBTP 22: Immunophenotyping of human pluripotent stem cells (PSCs) using flow cytometry

Miltenyi Biotec-tested panel 22 (MBTP 22)

This application protocol describes the quantitative analysis of human pluripotent stem cell (PSC) differentiation potential using flow cytometry. Human PSCs can easily be differentiated using the StemMACS™ Trilineage Differentiation Kit which enables directed and reproducible differentiation into ectodermal, mesodermal, and endodermal lineages ready for downstream applications, such as flow cytometric analysis.

Protocol

Gating strategy showing the analysis of human PSCs. Human PSCs were stained using the previously described panel to analyze their differentiation potential. Exclusion of debris using SSC-A/FSC-A gating was initially performed on meso-, ecto-, and endoderm samples, respectively (A, B, C). Propidium iodide was added to mesoderm samples shortly before flow cytometric analysis for exclusion of dead cells (D). No dead cell exclusion was done for endo- and ectoderm samples, since these cells were fixed. For mesoderm samples CD140b+ and CD144+ cells were gated (E), for ectoderm Sox2+ PAX-6+ cells (F), and for endoderm Sox17+ CD184 (CXCR4)+ cells (G).

Materials

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