Fig. 5
Immobilised Delta-Like 1 induces the same lineage skewing of hematopoietic progenitor cells in stirred bioreactor systems. CD34+ enriched cells (95% CD34+) were cultured with iDL1, alongside untreated, soluble DL1 and blank particles as controls, in a mechanically agitated suspension bioreactor (n = 3). a As previously shown in the static platform, 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 node defined phenotypically as CD34hiCD38negCD90negCD133hiCD135hi. b The addition of iDL1 to CD34+ enriched cells in a stirred culture system led to the same trend in lineage skewing previously identified in static. Evaluation of the percentage abundance within the progenitor node at day 12 identified high levels of iDL1 resulted in significantly more progenitor cells compared to all controls. c Cluster tree of CD15 expression, the parent CD15hi node was identified. d Evaluation of percentage abundance within the parent CD15hi node at day 12 identified, as in the static platform, culture with high levels of iDL1 resulted in significantly less mature CD15hi cells compared to all controls. Values were considered (*) statistically significant for P < 0.05