Fig. 4
Immobilised Delta-Like 1 can induce skewed lineage output of hematopoietic progenitor cells in static culture. CD34+ enriched cells (95% CD34+) were cultured with iDL1 at three concentrations alongside three controls: untreated, soluble DL1 and blank particles only (n = 5). a FCS files were uploaded into Cytobank and CITRUS cluster trees derived, clustering on CD34, CD33, CD38, CD45ra, CD90, CD133 and CD135 marker intensities. Cluster trees of CD34, CD38, CD90, CD133 and CD135 are shown, identifying the progenitor nodes defined phenotypically as CD34hiCD38negCD90negCD133hiCD135hi. b Progenitor nodes (identified in 4A) were evaluated for their percentage abundance at day 6 (i) and day 12 (ii), cells cultured with high levels of iDL1 contained a significantly higher percentage of progenitor cells compared to all controls. All concentrations of iDL1 significantly improved progenitor abundance compared with untreated and soluble DL1 controls at day 12. Further phenotypic analysis was undertaken at day 12 to evaluate the expression of more mature surface markers. FCS files were uploaded into Cytobank and CITRUS cluster trees derived, clustering on CD13, CD14, CD15, CD24, CD33, CD38 and CD123 marker intensities. c Cluster tree of CD14 expression, the parent CD14hi node was identified. d Evaluation of percentage abundance within the parent CD14hi node at day 12 identified culture with high and medium levels of iDL1 resulted in significantly less mature CD14hi cells compared to all controls. e Cluster tree of CD15 expression, the parent CD15hi node was identified. (F) Evaluation of percentage abundance within the parent CD15hi node at day 12 identified culture with high levels of iDL1 resulted in significantly less mature CD15hi cells compared to all controls. All concentrations of iDL1 significantly decreased CD15hi abundance compared with untreated and soluble DL1 controls. Values were considered (*) statistically significant for P < 0.05, (**) very statistically significant for P < 0.01