F. Rinaldi, M. Rynning, D. Galitz, R. Fuerstenberg, L. Leong and J. Aho
The ability of pluripotent stem cells to differentiate into any tissue of the body has the potential to revolutionize medicine. To fully realize this potential, robust and standardized differentiation and characterization protocols are necessary. In this study, we describe the use of Luminex® multi-analyte technology for non-invasively characterizing stem cells undergoing differentiation as well as for optimizing individualized differentiation protocols. Luminex® technology allows for the simultaneous quantification of up to 100 proteins within a single small-volume sample. We utilized the multi-analyte screening power of Luminex® assays to profile the levels of cytokines and growth factors in cell culture media at key stages during the differentiation of pluripotent stem cells into hepatocytes. Cytokine and growth factor expression profiles were obtained from human induced pluripotent stem (hiPS) and human embryonic stem (hES) cell lines with known differences in hepatocyte 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 albumin expression. We hypothesize that the multi-analyte profile of cell lines with robust differentiation efficiency will differ from cell lines with lower efficiencies. Using Luminex® multi-analyte technology we will be able to identify particular analytes that are predictive of differentiation success. Additionally, this data can be used to identify alternate factors that enhance differentiation and/or maturation of the differentiated cells.