Extended Data Fig. 3: Optimization of ER + and HER2 + PDxO culture.
(a) Growth response of HCI-011 PDxO to culture medium additives, quantified as live cell area. Mean ± s.e.m. All conditions are n=3 biological replicates except A83-02 + Y-27632 and Y-27632+CHIR (3 days) n=5 biological replicates; Y-27632, A83-01, CHIR (3 days), SB-202190 n=6 biological replicates. (b) Effect of other common breast cancer medium supplements on growth of HCI-011 PDxOs, quantified by Cell Titer Glo 3D (CTG-3D) assay to measure ATP content. Mean ± s.e.m., n=3 biological replicates. (c) Effect of other common breast cancer medium supplements on growth of HCI-011 PDxOs, quantified by CTG-3D. Mean ± s.e.m., n=3 biological replicates. Statistical comparisons to control condition (minus all (left) or +Y-276362 (right)) by ordinary two-way ANOVA, uncorrected Fisher’s LSD with single pooled variance (d) HCI-011 PDxO organoid size (radius) after addition of Y-27632, NAC and FGF2 to PDxO cultures. Each gray dot represents one organoid. Data are shown as mean and individual data points: control n=449 individual organoids, bFGF n=313 individual organoids, bFGF + NAC n=344 individual organoids. Statistical comparisons to control (+Y-27632) by ordinary two-way ANOVA, uncorrected Fisher’s LSD with single pooled variance. (e) Comparison of culture doubling time of HCI-011 and HCI-017 when established in our optimized ER + PDxO media or organoid media published by Sachs et al., 2018. Mean ± s.e.m. HCI-017 n=6 biological replicates, HCI-011 Sachs et al n=4 biological replicates, HCI-011 PDxO n=3 biological replicates. Statistical comparisons to control condition (Sachs et al., 2018) within each line by ordinary two-way ANOVA, uncorrected Fisher’s LSD individual variances computed from each comparison. (f) Immunohistochemistry of HER2 showing variable HER2 staining in PDxOs and their parental PDX tumor with HER2 + histories. HCI-005 (top) and HCI-008 (bottom) are shown. Each IHC staining has been performed once for each model. Scale bar corresponds to 50 um. (g) Effect of additional medium supplements on growth of HCI-005 PDxOs, quantified by CTG-3D assay (left) and live cell area (right). Mean ± s.e.m., n=3 biological replicates. Statistical comparisons to control condition (+Y-27632+NAC + FGF2) by ordinary two-way ANOVA, uncorrected Fisher’s LSD with single pooled variance.
