Fig. 2

Exploratory morphological profiling HTS identifies growth factor combinations that induce distinct MSC phenotypes and enhance proliferation. (A) MSC proliferation of hits and controls after 48 h of culture where the dashed line represents initial seeding density. Cell count data from each media type was compared to SCM and CDBM on a per-plate basis via ordinary one-way ANOVA corrected for multiple comparisons via Dunnett’s test. *p < 0.05 compared to SCM, #p < 0.05 compared to CDBM. n = 12 per plate for SCM, n = 12 per plate for CDBM, n = 4 per plate for SCM+, n = 4 for each treatment group. (B) Hierarchical clustering of treatment groups that satisfy ‘hit’ criteria (i.e. significantly higher proliferation and distinct phenotype from controls based on significant mp-value (see Supplemental Figures)) distinguishes 8 hits from each other based on morphological principal components. (C) Table of final identified hits to be included in subsequent validation screen. (D) PCA plot representing morphological distinction between treatment groups and controls. (E) Representative images from screen of MSCs grown in SCM and various hit media (red = actin, Golgi, plasma membrane, blue = nucleus, green = endoplasmic reticulum, magenta = mitochondria. Scale bar = 200 μm)