Single cell sequencing analysis of control, photon- and proton- irradiated salivary gland organoids at day 7 and control salivary gland organoids at day 11

The specific impact of different radiation types, such as photons and protons, and their effects on normal stem cell activity and tissue regeneration are unknown. In this study, we used a scRNA-seq approach to characterize our mouse salivary gland organoid model at day 7 and 11 and assess the effects of photon and proton irradiation on salivary gland stem/progenitor cells at day 7. Mouse salivary gland organoids derived from 4 different animals were irradiated at day 5 with either 7 Gy photons or 7 Gy protons (plateau). Non-irradiated (control), photon- and proton-irradiated organoids were collected at day 7 (2 days after irradiation) and pooled before the sequencing run. After trypsinization, propidium iodide (PI) was used to isolate live cells (PI negative) by flowrescence-activated cell sorting (FACS). Cells were then analysed using scRNA-seq. Fresh isolated submandibular salivary gland tissue was also included in the analysis together with non-irradiated 11-day old salivary gland organoids. 11-day salivary gland organoids were run separately.