Fig. 3: CRISPR-modification of oviductal and OSE organoids.

a Strategy to generate the mutant lines using CRISPR-Cas9. The lines were trypsinized to single-cell suspension followed by sgRNA transfection. Three days after transfection Nutlin-3a was added to the medium to allow mutant Trp53 organoid outgrowth. In 2 weeks emerging clonal organoids were picked, expanded and screened. Scale bar, 200 μm. b Hypothesized tumor progression model of HG-SOC and chosen genes to build comparable progression models. STIC—serous tubal intraepithelial carcinoma. c Brightfield images of comparable clones with the different set of mutations from both oviductal and OSE origin. Scale bar, 100 μm. d Immunohistochemical stainings for H&E to visualize the more detailed phenotype of oviductal and OSE clones. Arrow heads point to the cellular stratification. Scale bar, 50 μm. c, d Org.—organoids. e, f Scatter plots presenting chromosome number distribution and mean of e oviduct wild-type and mutant clones or f OSE wild-type and mutant clones, based on organoid metaphase spreads (n = 20 spreads per biologically independent clones). c–f OSE—ovarian surface epithelium, WT—wild-type, T—Trp53 mutant; TB—Trp53, Brca1 mutant; TBN—Trp53, Brca1, Nf1 mutant; TBP—Trp53, Brca1, Pten mutant.