ANN identifies HSC-like cells in iPSC-derived hematopoietic cells. (A) Schematic of the ANN architecture for label transfer composed of an input layer (3479 units), 2 fully connected hidden layers (64 and 32 units), and a 28-unit softmax layer corresponding to cell types in the source domain (human fetal liver scRNAseq data) used for training. Classification of cell types in the target domain of human iPSC-derived single-cell transcriptomes (test data). (B) Proportion of cells labeled HSC/MPP at day 10 or day 13 of hiPSC differentiation in vitro. (C) Proportion of in vitro–derived CD235a⁻ progenitors and CD235a⁻CD43⁺ cells labeled HSC/MPP by the ANN (ND, not detected). (D) UMAP visualization of the integrated data set containing in vivo–derived and in vitro–annotated HSCs/MPPs. (E) Heatmap of differentially expressed genes coding for transcription factors obtained from the comparison of in vivo– and in vitro–derived HSCs/MPPs. (F) Transcription factor binding motifs enriched upstream of the 54 genes identified as differentially expressed between HSC/MPP generated in vitro and in vivo. UMAP, uniform manifold approximation and projection.