Predicting Cell Line Variability in Cardiomyocyte Differentiation Efficiency Using Non-invasive Multi-analyte Luminex Assays

F. Rinaldi, M. Freeman, M. Rynning, R. Fuerstenberg, and J. Aho


The ability of pluripotent stem cells to differentiate into any tissue of the body has the potential to revolutionize medicine. One technical challenge that must be overcome to fully realize this potential is generating patient-derived pluripotent stem cells that can efficiently and robustly differentiate into specific tissues. Even using standardized differentiation protocols, differentiation efficiency is highly variable across cell lines from both human embryonic stem (hES) and human induced pluripotent stem (hiPS) cells. In this study, we demonstrate the power of Luminex® Assays as an early detection method to assess pluripotent cell line variability and to determine cell line differentiation efficiency. Various hES and hiPS cell lines were differentiated into cardiomyocytes using the standardized protocol and reagents in the StemXVivo® Cardiomyocyte Differentiation Kit. Combining our standardized differentiation protocol with multi-analyte Luminex® assays enabled us to profile the changes in cytokine and growth factor levels in cell culture media at key stages during the differentiation. We found that cytokine and growth factor expression profiles varied across hiPS cell lines with known differences in cardiomyocyte differentiation efficiency. Because analytic samples are obtained from culture media, the cells are able to continue through the differentiation process and be analyzed for efficiency by assessing beating and cardiac-specific marker expression via immunocytochemistry. We demonstrate that the multi-analyte profile of cell lines with robust differentiation differs from cell lines with lower efficiencies. Using Luminex® multi-analyte technology we identified particular analytes that are predictive of differentiation success. Additionally, this information can be used for identification of important pathways involved in stem cell differentiation and/or maturation into cardiomyocytes.