(A) ELISA assay for mouse TPO stability in RPMI-based culture media after 24 hours. Mean ± S.D of technical duplicates from biological replicates.
(B) ) Schematic summary of TPO-dependent mouse 32D/MPL cells grown in various RPMI-based medias supplemented 10ng/ml TPO.
(C) Total numbers of 32D/MPL cells after a 14-day culture in the RMPI-based medias described in (B) with TPO at 10 ng/ml. Dotted line indicates starting cell concentration. Mean ± S.D of four independent cultures.
(D) Total numbers of primary human AML cells (two AML samples from the Stanford Hematology Biobank) after a 14-day culture in MEMa-based medias, with indicated supplements and cytokines (TPO, SCF, FLT3L, IL-6, IL-3, GM-CSF, and G-CSF; all 20 ng/ml). Dotted line indicates starting cell concentration. Mean ± S.D of four independent cultures.
(E) Total numbers of human K-562 cells over a 14-day culture in the RMPI-based medias with indicated supplements. Mean of four independent cultures.
In all panels, statistical analysis was performed by one-way ANOVA (analysis measured against the PVA+ITSX condition with significance indicated by: *, p<0.05, **, p<0.01, ***, p>0.001; ****, p>0.0001; n.s, non-significant.