Proteomics-based dissection of human endoderm progenitors by differential cell capture on antibody array.

Molecular & Cellular Proteomics : MCP
Revital SharivkinYoav Soen

Abstract

Heterogeneity, shortage of material, and lack of progenitor-specific cell surface markers are major obstacles to elucidating the mechanisms underlying developmental processes. Here we report a proteomics platform that alleviates these difficulties and demonstrate its effectiveness in fractionating heterogeneous cultures of early endoderm derived from human embryonic stem cells. The approach, designated differential cell-capture antibody array, is based on highly parallel, comparative screening of live cell populations using hundreds of antibodies directed against cell-surface antigens. We used this platform to fractionate the hitherto unresolved early endoderm compartment of CXCR4+ cells and identify several endoderm (CD61+ and CD63+) and non-endoderm (CD271+, CD49F+, CD44+ and B2M+) sub-populations. We provide evidence that one of these sub-populations, CD61+, is directly derived from CXCR4+ cells, displays characteristic kinetics of emergence, and exhibits a distinct gene expression profile. The results demonstrate the potential of the cell-capture antibody array as a powerful proteomics tool for detailed dissection of heterogeneous cellular systems.

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Citations

Aug 31, 2013·Omics : a Journal of Integrative Biology·Aihua ZhangXijun Wang
Mar 7, 2018·Clinical Proteomics·Ziqing ChenSheng-Ce Tao
Jul 13, 2014·The Journal of Biological Chemistry·Keren Bahar HalpernMichael D Walker
Jul 28, 2020·Cellular and Molecular Life Sciences : CMLS·Anna MeyfourGhasem Hosseini Salekdeh

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