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. 2018 Jul 12;3(13):e95319. doi: 10.1172/jci.insight.95319

Figure 5. Increased cell death and pyroptosis in old hematopoietic progenitors.

Figure 5

(A) Frequency of dead cells among TLPs (using LIVE/DEAD staining assay) at 7, 14, 21, and 28 days upon in vitro differentiation of FACS-isolated CD34+ HPCs from young (Y, n = 9) or old (O, n = 10) healthy adults in the OP9-DL1 coculture system. (B) Representative image and staining levels for annexin V and PI in bead-enriched CD34+ cells from young (n = 5) or old adult (n = 5) PBMCs. (C) Increased mRNA expression of the proapoptotic gene P2X7 in FACS-isolated HPCs from old (n = 10) healthy adults compared with HPCs from young (n = 7) subjects. (D) Representative staining and expression levels of cleaved caspase 1 in CD34+ cells from young (n = 5) or old (n = 5) adult PBMCs. Data are expressed as ratio of stimulated versus unstimulated cells. Bars indicate the median. (E) Enhancement of in vitro survival of TLPs at 14 or 21 days in the presence of PPAD (P2X7 inhibitor, at 20 μM) or VX-765 (caspase 1 inhibitor, at 0.1 μM) from day 7 in the OP9-DL1 culture system. Survival is expressed as the ratio of dying cells at day 14 or 21 over day 7 in culture (n = 5 in the presence of PPAD or VX-765 and n = 10 in the absence of inhibitors). Columns indicate mean (+SEM). The Mann-Whitney test was used for comparisons.