Fig. 1

Establishment of a hESC-to-HSPC differentiation system and the application of GelMA in vitro culture. a Schematic diagram of the differentiation from hESCs to HSPCs. b Representative morphology of hESCs-H9 cells (scale bar, 100 µm). c Representative morphologies of HSPCs induced from hESCs-H9 cells (scale bar, 50 µm). d Flow cytometry analysis of the expression of CD34, CD43, and CD45 on the induced HSPCs. e Immunofluorescence staining for the detection of CD34, CD43, and CD45 expression on the induced HSPCs (scale bar, 20 µm). f Schematic diagram of 2D and 3D-GelMA culture. g Percentage of CD34⁺ cells on day 7 for each condition. Results are presented as the mean ± SD from three independent experiments. Unpaired Student’s t-test, ***p < 0.001. h Representative morphologies of different colony types (CFU-GEMM, CFU-GM and BFU-E) and total number of colonies under different culture conditions (scale bar, 100 µm). Results are presented as the mean ± SD from three independent experiments. Unpaired Student’s t-test, **p < 0.01. i Percentage of CD34⁺ and CD45⁺ cells in G0/G1, S and G2/M phase of the cell cycle for each condition after 7-day culture. Results are presented as the mean ± SD from three independent experiments. Two-way ANOVA, ***p < 0.001. j Intracellular ROS levels within CD34⁺ cells were determined utilizing DCFH assay via flow cytometry. Results are presented as the mean ± SD from three independent experiments. Unpaired Student’s t-test, **p < 0.01