Fig. 1

Co-culture with MSCs rescues the busulfan-induced damage of small intestine organoids. A Schematic overview of the in vitro co-culture model of MSCs and small intestine organoids damaged by treatment with busulfan. Single cell small intestine organoids were embedded in matrigel and grown for 5 days (day 0–5) and treated with busulfan (35 µM) for 48 h (day 5–7). Organoids damaged by treatment with busulfan or control organoids were co-cultured with 5000, 10,000, and 50,000 MSCs for 24 h or 48 h (day 7–8/9) and the surface area of organoids was measured. B Representative images of control organoids and organoids treated with busulfan co-cultured without or with 10,000 MSCs at 48 h after co-culture are shown. C Busulfan reduced the size of the organoids in organoid donor 1 and organoid donor 2. D Co-culture with MSCs increased the size of the organoids after treatment with 5000, 10,000, and 50,000 cells in organoid donor 1 and increased the size of the organoids after treatment with 50,000 MSCs in organoid donor 2. E Co-culture with 5 out of 9 tested bone marrow derived-MSC donors increased the size of organoids treated with busulfan. The quantification of surface area of the organoids was represented as fold change as compared to control. Results are shown as means ± SEM of data from 2 different organoid donors co-cultured with at least 3 MSC donors. Due to the large biological variation in organoid size, the statistical analysis of the effect of individual MSC donors on the size of busulfan-induced damaged organoids (E) was based on all evaluable individual organoids (of at least 3 organoid/matrigel droplets cultured in different wells). Scale bars, 1000 µm. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 as compared to control (Kruskal–Wallis test or a Mann Whitney test)