Establishment, Characterization, and Tissue Comparison of Canine Pancreatic and Endometrial Organoids, Alongside Patient Matched Organoids

Organoids are 3-dimensional (3D) stem cell-derived cultures that offer a variety of technical advantages compared to traditional 2-dimensional (2D) cell cultures. Although murine models have proved useful in biomedical research, rodent models often fail to adequately mimic human physiology and disease progression, resulting in poor preclinical prediction of therapeutic drug efficacy and toxicity. An interesting alternative is to use the canine model in research, due to its numerous similarities to humans (shared environment, intact immune system, and development of civilization diseases). The use of canine organoids in drug testing and disease modeling has been limited by the number of models as well as the depth of characterization. Here, we report the establishment, maintenance, and molecular characterization of six adult-stem cell-derived canine organoid cell lines from endometrium, pancreas, urinary bladder, kidney, lung, and liver from two genetically related canines. The organoids of the canine endometrium and pancreas have not yet been described in the literature. Furthermore, scRNASeq was utilized on a subset of the organoids to identify organoid specific transcriptomic signatures including lung (SFTPC), pancreas (TFF3), kidney (GJB2), and bladder (UPK1A). In total, six tissues and organoid lines from each donor were characterized, allowing for a unique, multi-organ comparison between these two individuals and identification of specific cell types within the organoids. scRNA-seq profiling of four healthy canine organoid lines.