Journal: bioRxiv
Article Title: Developmental regulation of endothelial-to-hematopoietic transition from induced pluripotent stem cells
doi: 10.1101/2024.09.24.612755
Figure Lengend Snippet: ( A ) HE population (cluster 75 in integrated dataset) used for differential gene expression analysis. ( B,C ) Gene ontology (GO) terms significantly upregulated (B) or downregulated (C) in iPSC-derived vs. HSC-competent human embryo HE (Metascape, q < 0.05). ( D ) Arterial endothelium (AE; cluster 36, red) and HE (cluster 75, blue) populations used for differential expression analysis. ( E,F ) GO terms significantly upregulated (E) or downregulated (F) during AE-to-HE transition (Metascape batch mode, q < 0.05). ( G ) HE (cluster 75, red) and hematopoietic stem and progenitor cell (HSPC; cluster 74, blue) populations used for differential expression analysis. ( H,I ) GO terms significantly upregulated (H) or downregulated (I) during HE-to-HSPC transition (Metascape batch mode, q < 0.05). ( J ) Expression of representative endothelial identity genes FLT1 and KDR in HSC-competent human embryos (CS14-17; red) and day 8 iPS (blue) AE (cluster 36), HE (cluster 75) and HSPCs (cluster 74). ( K ) Expression of HSC genes HLF , HOXA9 , MECOM , and SPINK2 in HSC-competent human embryos (CS14-17; red) and day 8 iPS (blue) AE (cluster 36), HE (cluster 75) and HSPCs (cluster 74). For (J, K) *q < 0.05, **q < 0.005, and ***q < 0.0005 using a linear regression fit to a mixed-negative binomial model accounting for batch effects.
Article Snippet: They were resuspended in EB culture media supplemented with final concentrations of 100 ng/mL human stem cell factor (hSCF, Peprotech #300-07), 100 ng/mL fms-like tyrosine kinase 3 ligand (hFLT3L, Peprotech #300-19), 50 ng/mL bone morphogenic protein 4 (hBMP4, R&D Systems #314-BP), 10 ng/mL granulocyte colony-stimulating factor (hG-CSF, Peprotech #300-23), 25 ng/mL interleukin-6 (hIL-6, Peprotech #200-06) and 10 ng/mL interleukin-3 (hIL-3, Peprotech #200-03) and again put on a shaker at 37°C.
Techniques: Expressing, Derivative Assay