• Document: Resolving Stem Cell Heterogeneity Using Flow Cytometry
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Resolving Stem Cell Heterogeneity Using Flow Cytometry Mirko Corselli, PhD Senior Scientist bdbiosciences.com We have not fully utilized the power of flow cytometry to address biological questions in other systems Flow cytometry is essential for our understanding of the hematopoietic system bdbiosciences.com Challenges in MSC Research Phenotypic characterization: • Heterogeneity • Lack of biomarkers associated with MSC function Functional characterization: • Need for standardized potency assays bdbiosciences.com Overview • Minimal criteria to define MSCs • Multicolor panels to identify MSC subpopulations • Surface marker screening to define new stem cell signatures • Multi-parametric assay to study MSC immunomodulatory activity bdbiosciences.com • MSCs must be plastic adherent. • MSCs must express CD73, CD105, and CD90 and lack expression of CD45, CD34, CD14, CD11b, CD79a, and CD19. • MSCs must differentiate into osteoblasts, adipocytes, and chondrocytes in vitro. bdbiosciences.com Analysis of MSCs by Flow Cytometry BM-MSCs Relative Cell Number Relative Cell Number Relative Cell Number Relative Cell Number 0.655 100 100 100 2 3 4 5 2 3 4 5 2 3 4 5 2 3 4 5 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 Lin PE CD90 FITC CD73 APC CD105 PerCP-Cy™5.5 ADSCs Relative Cell Number Relative Cell Number Relative Cell Number Relative Cell Number 0.476 99.9 99.9 99.8 2 3 4 5 2 3 4 5 2 3 4 5 2 3 4 5 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 Lin PE CD90 FITC CD73 APC CD105 PerCP-Cy™5.5 BD Stemflow™ MSC Analysis Kit bdbiosciences.com

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