- Human pluripotent stem cells can be differentiated into a variety of different cell types, for instance cardiomyocytes. Especially in the context of future application in regenerative medicine, it is crucial to understand the developmental processes taking place upon differentiation. Moreover, the identification of a panel of cell surface markers suitable for characterization and purification is necessary to ensure quality of human pluripotent stem-cell derived products. In this study, we used quantitative mass spectrometry to characterize proteomic changes in early mesendodermal differentiation. Two human pluripotent stem cell lines, one embryonic (H3) and one induced pluripotent stem cell line (I2), were analyzed under pluripotent conditions and after two days of embryoid body-based differentiation. Functional clustering and enrichment analysis showed down-regulation of proteins associated with pluripotency and the tricarboxylic acid cycle at day two. In contrast, proteins related toHuman pluripotent stem cells can be differentiated into a variety of different cell types, for instance cardiomyocytes. Especially in the context of future application in regenerative medicine, it is crucial to understand the developmental processes taking place upon differentiation. Moreover, the identification of a panel of cell surface markers suitable for characterization and purification is necessary to ensure quality of human pluripotent stem-cell derived products. In this study, we used quantitative mass spectrometry to characterize proteomic changes in early mesendodermal differentiation. Two human pluripotent stem cell lines, one embryonic (H3) and one induced pluripotent stem cell line (I2), were analyzed under pluripotent conditions and after two days of embryoid body-based differentiation. Functional clustering and enrichment analysis showed down-regulation of proteins associated with pluripotency and the tricarboxylic acid cycle at day two. In contrast, proteins related to the proteasome and annexin family were up-regulated upon differentiation. Among the proteins that were down-regulated upon differentiation in both, H3 and I2, the membrane protein junctional adhesion molecule A (JAM-A) emerged as potentially associated with pluripotency. Flow cytometry and immunocytochemistry further confirmed down-regulation of JAM-A on the cell surface of human induced pluripotent stem cells that were differentiated toward mesendoderm for just two days.…
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