Figure 2: EDICTS captures the evolution of stem cell lineage restriction in time, as evidenced by four dimensional spatiotemporal image informatics of H3K4K27me3 expression in developing hMSCs.

(A) Cells were either maintained in a basal stem state or induced to differentiate toward adipogenic and osteogenic lineages. hMSCs were subsequently fixed, immunolabeled and imaged for H3K4me3 (green) and H3K27me3 (red) at 9 time points over two weeks. All images shown are 30 μm × 30 μm. (B) Principal component analysis (PCA) of their texture descriptors reveals the ability to classify the three conditions as early as 72 hours post seeding, with classification statistical accuracy improving over each time point. An average sensitivity/specificity value above 80% indicates high classification accuracy, which is achieved at the 72 hour mark between all cell types, and as early as between 24 and 48 hours between undifferentiated hMSCs and hMSCs induced to differentiate. The distances between the centroids of each cell phenotype steadily increases over time, with greater separation between undifferentiated hMSCs and either differentiated cell type (i.e. adipogenic and osteogenic) compared to adipogenic versus osteogenic, suggesting that (i) these texture descriptors directly correlate to “degrees” of phenotypic commitment, and (ii) generally distinguish undifferentiated cells from differentiated cells earlier than distinguishing different differentiated cell types from each other.