TY - JOUR T1 - RNA-sequencing of single cholangiocyte-derived organoids reveals high organoid-to organoid variability JF - Life Science Alliance JO - Life Sci. Alliance DO - 10.26508/lsa.202101340 VL - 5 IS - 12 SP - e202101340 AU - Kristin Gehling AU - Swati Parekh AU - Farina Schneider AU - Marcel Kirchner AU - Vangelis Kondylis AU - Chrysa Nikopoulou AU - Peter Tessarz Y1 - 2022/12/01 UR - https://www.life-science-alliance.org/content/5/12/e202101340.abstract N2 - Over the last decades, organoids have been established from most of the tissue-resident stem and iPS cells. They hold great promise for our understanding of mammalian organ development, but also for the study of disease or even personalised medicine. In recent years, several reports hinted at intraculture organoid variability, but a systematic analysis of such heterogeneity has not been performed before. Here, we used RNA-seq of individual intrahepatic cholangiocyte organoids to address this question. We find that batch-to-batch variation is very low, whereas passage number has a profound impact on gene expression profiles. On the other hand, there is organoid-to-organoid variability within a culture. Using differential gene expression, we did not identify specific pathways that drive this variability, pointing towards possible effects of the microenvironment within the culture condition. Taken together, our study provides a framework for organoid researchers to properly consider experimental design. ER -