Fig. 5: Impact of HC/MΦ and MC on BMEC in WBM culture.

a–d P0 WBM cultures were divided in three conditions: control and supplement at day 0 with additional HC/Mɸ or MC. a, b Snapshot images at day 14 culture showing tdT⁺ BMECs in P0 WBM cultures that were supplemented with a HC/MΦ or b MC, compared to control; two representative fields; Scale bar, 2000 μm. n = 12. c Fold change in BMEC numbers: tdT⁺ (left), Emcn⁻ (middle) and Emcn⁺ (right) when P0 WBM culture is supplemented with HC/Mɸ or MC, vs. culture control. Data are presented as the mean ± SEM. Emcn⁺ cells expanded on average 48-fold in control cultures (set up in this graph at value 1), 96-fold when supplemented with HC/MΦ, and 24-fold when supplemented with MC. BMEC + MΦ, n = 8; BMEC + MC, n = 6. n = 3 independent experiments. Statistics for all comparisons shown were determined using unpaired t-test, two-sided. tdT⁺ MΦ, p = 0.0026; tdT⁺ MC, p < 0.0001; Emcn⁻ MΦ, p < 0.0001; Emcn⁻ MC, p < 0.0001; Emcn⁺ MΦ, p < 0.0001; Emcn⁺ MC, ***p < 0.001, and ****p < 0.0001. Source data are provided as a Source Data file. d Total cell number of Emcn⁺ and Emcn⁻ cells of P0 WBM culture with or without additional HC/MΦ or MC at different WBM cell densities. Representative of n = 3 independent experiments. Source data are provided as a Source Data file. e, f P1 WBM cultures were supplemented with additional HC/MΦ and cultured for seven days: e total cell number and f fold changes of Emcn⁺ and Emcn⁻ cells supplemented with additional HC/MΦ vs. control, at different cell densities; n = 3. Data are presented as the mean ± SEM. Statistics for all comparisons shown were determined using unpaired t-test, two-sided. e Cell density 1, Emcn⁻, p = 0.0020; cell density 1/4, Emcn⁻, p = 0.0006; cell density 1/8, Emcn⁻, p = 0.0001; Emcn⁺ p = ns at all cell densities. f Cell density 1, Emcn⁻, p = 0.0006; cell density 1/4, Emcn⁻, p = 0.0004; cell density 1/8, Emcn⁻, p < 0.0001; Emcn⁺ p = ns at all cell densities. **p < 0.01, ***p < 0.001, and ****p < 0.0001. Source data are provided as a Source Data file. g P0 WBM cultures were supplemented with recombinant M-CSF (50 ng/mL) and cultured for fifteen days. Total cell number of Emcn⁺ and Emcn⁻ cells of P0 WBM culture with or without M-CSF; n = 3 independent cultures. Data are presented as the mean ± SEM. Statistics were determined using unpaired t-test, two-sided *p = 0.0336.