Journal: Frontiers in Immunology

Article Title: Ex vivo- generated lymphoid progenitors encompass both T cell and innate lymphoid cell fates

doi: 10.3389/fimmu.2025.1617707

Figure Lengend Snippet: Activation of BCL11B regulatory elements does not restrict NK cell potential. (A) Experimental design for assessing BCL11B role in human T cell commitment. EGFP-BAC reporter was integrated at exon 1 of endogenous BCL11B gene in hPSC, resulting in a monoallelic disruption of this gene and the creation of BCL11B -EGFP reporter cell line. BCL11B -EGFP hPSCs were differentiated into hematopoietic progenitors (HP) for 9 days. The generated HP cells were isolated for early T cell differentiation on OP9-DLL4 for 14 more days. At this time, T9+T14, BCL11B -EGFP negative and positive progenitor cells had been sorted by FACS and secondary cultures were performed to assess their myeloid, NK and T cell potential. Phenotypes were assessed at each stage by flow cytometry for myeloid, lymphoid, NK or T membrane markers. (B) Representative dot plot of flow cytometry analysis of BCL11B -EGFP hPSC-derived progenitors upon sorting at day T9+T14 after culture on OP9-DLL4 for 14 days. At this stage, CD45 + CD56 - CD7 + cells are clearly distributed into two populations, according to BCL11B -EGFP expression. (C, D) BCL11B-EGFP neg/+ progenitors were co-cultured for 10 days with OP9-DLL4 with IL15 for NK differentiation. (C) Representative contour plots of flow cytometry analysis illustrating the expression of CD56, delineating NK differentiation, and CD16, translating NK maturation. (D) Bar plot representing the mean ± SD of CD56-expressing NK-primed cells in three independent experiments. *p<0.05 values are for paired Student’s t-test. (E, F) BCL11B -EGFP neg/+ progenitors were co-cultured 5 days with OP9 for myeloid differentiation. (E) Representative contour plots of flow cytometry analysis illustrating CD11b expression translating myeloid priming. (F) Bar plot representing the mean ± SD of CD11b-expressing myeloid progenitors in three independent experiments. *p<0.05 values are for paired Student’s t-test. (G, H) BCL11B -EGFP neg/+ progenitors were co-cultured 7 days with OP9-DLL4 devoid of IL15 for T cell differentiation. (G) Representative contour plots of flow cytometry analysis illustrating the expression of CD7 and CD5 translating T cell differentiation. (F) Bar plot representing the mean ± SD of CD7 and CD5-expressing T cell progenitor cells in four independent experiments.

Article Snippet: 209Bi , CD11b (Mac-1) , Fluidigm , ICRF44 , 3209003C.

Techniques: Activation Assay, Disruption, Generated, Isolation, Cell Differentiation, Flow Cytometry, Membrane, Derivative Assay, Expressing, Cell Culture