Figure 1: Single-cell transcriptional analysis identifies a subpopulation of human ASCs with putatively enhanced regenerative potential.

(a) Single-cell transcriptional screening of all known cell SMs to identify those with differential expression (most useful for cell subtyping). Gene expression presented as fold change from median (yellow—high expression, 32-fold above median to blue—low expression, 32-fold below median; grey—no expression). (b) Single-cell analysis focused on high copy number, differentially distributed SM genes identified a cell subpopulation present across repeated k-means clusterings. (c) Linear discriminate analysis (LDA) identified SMs for prospective subpopulation isolation, with ROC analysis of cluster sensitivity and specificity utilizing the ‘best’ individual or groups of genes determined using forward feature selection. (d) Single-cell confirmation of prospective hASC subpopulation isolation via FACS using two LDA-defined SMs (DPP4 and CD55). (e) Positive hASC subpopulation enrichment enhances gene expression distributions for multiple genes related to tissue regeneration (selected significantly affected genes displayed as determined via Kolmogorov–Smirnov testing). (f) Single-cell whisker plots and pooled cell RT-PCR demonstrating a confirmation of selected single-cell gene distribution findings on a population level. (g) Top scoring IPA-constructed transcriptome network based on the genes significantly increased following positive hASC selection. Significant ‘seed’ genes are coloured in red to distinguish them from the remaining ‘inferred’ entities in the network. *indicates P≤0.05 for positive selection versus hASCs or negative selection, via one-way ANOVA. Error bars represent s.e.m.