Fig. 5

TNFα enhances the in vitro and in vivo T-cell potential of CD7⁺ HTLPs derived from CB or mPB HSPCs. Graphs showing the mean frequencies of CD4⁺CD8⁺ cells (A) and CD3⁺ cells (B) obtained after 1, 2, 3 and 4 weeks of coculture of day-7 CB or mPB HSPC-derived HTLPs (with or without TNFα treatment) with OP9-hDL1 stromal cells (mean ± SEM, n = 3). The p values were calculated using a paired two-tailed t test: *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001. C Analyses of TCRδ, TCRγ and TCRβ rearrangements in T cells differentiated from CB or mPB CD7⁺ HTLPs after OP9-hDL1 coculture. Each peak represents the fluorescence intensity of the corresponding rearrangement loci; positive control: peripheral blood lymphocytes (PBLs). D Representative photographs of the thymus four weeks after intrahepatic injection of 5 × 10⁵ CB or mPB HSPC-derived HTLPs (cultured with or without TNFα treatment) into 1- to 4-day-old NSG mice. Graphs showing the chimerism and numbers of hCD45⁺ cells in the thymus of CB (E) and mPB (F) HSPC-derived HTLP recipients. Graphs showing the frequencies and numbers of CD4⁺CD8⁺ cells for CB-HTLP (G) and mPB-HTLP (H). Graphs showing the frequencies and numbers of CD3⁺ cells for CB-HTLP (I) and mPB-HTLP (J). Each dot represents a recipient mouse. The p values were calculated by one^-way RM ANOVA: *p ≤ 0.05; **p ≤ 0.01