Journal: Angiogenesis
Article Title: High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins
doi: 10.1007/s10456-025-09985-5
Figure Lengend Snippet: Abnormal proliferation of KO cells in mosaic blood vessel organoids and EC co-cultures. A In mosaic blood vessel organoids consisting of CCM1 KO and wild-type cells (= CCM1 KO/WT) or CCM3 KO and wild-type cells (= CCM3 KO/WT), abnormally increased proliferation of CCM1 KO and CCM3 KO cells was observed. In mosaic blood vessel organoids consisting of CCM2 KO and wild-type cells (= CCM2 KO/WT), a reduced proliferation of CCM2 KO cells was found. KO cells were labeled with mEGFP. WT cells were labeled either with mTagRFPT or mEGFP (scale bar: 200 µm). B Mean mEGFP intensities in mosaic organoids consisting of mEGFP-labeled KO and mTagRFPT-labeled WT cells (= KO/WT) were normalized to the mean mEGFP intensity in control mosaic organoids consisting of mEGFP-labeled WT and mTagRFPT-labeled WT cells (= WT/WT). Mosaic blood vessel organoids were differentiated in three independent runs (n = 38–48 per genotype). Data are presented as individual data points and means. Statistical significance was assessed using the Mann–Whitney U-test with Welch's correction (** P < 0.01, *** P < 0.001). C , D In a 2D co-culture validation approach, mEGFP-labeled CCM1 or CCM3 KO iECs and mTagRFPT-labeled WT iECs were seeded in a 1:9 ratio (= CCM1 KO/WT or CCM3 KO/WT) and cultivated in either EndoGRO-MV (C) or STEMdiff EC expansion medium (D). Co-cultures of mEGFP-labeled WT and mTagRFPT-labeled WT iECs (= WT/WT) served as controls (scale bar: 200 µm). E , F After 6 days, the area of mEGFP-labeled cells compared to all cells was determined using FIJI software (n = 4 per genotype in three independent biological replicates). KO/WT = co-cultures of mEGFP-labeled KO and mTagRFPT-labeled WT iECs; WT/WT = co-cultures of mEGFP-labeled WT and mTagRFPT-labeled WT iECs. Data are presented as individual data points and means. Multiple two-sample t-tests with Welch's correction and Holm-Šídák adjustment for multiple testing were used for statistical analyses (* = Padj < 0.05; ** = Padj < 0.01)
Article Snippet: CCM1 KO AICS-0016 hiPSCs, CCM3 KO AICS-0016 hiPSCs, WT AICS-0016 hiPSCs, and WT AICS-0054 hiPSCs were differentiated into ECs (iECs) using the STEMdiff Endothelial Differentiation Kit (Stemcell Technologies) as previously described [ ].
Techniques: Labeling, Control, MANN-WHITNEY, Co-Culture Assay, Biomarker Discovery, Software
