Establishment and Transcriptomic Characterization of Canine Organoids from Multiple Tissues

Christopher Zdyrski^1,2*Vojtech Gabriel³Oscar Ospina⁴Hannah Nicholson¹Michael Catucci¹Bryan J Melvin¹Hannah Wickham³Dipak Kumar Sahoo³Kimberly Dao²Leeann S. Aguilar Meza³Abigail Ralston²Leila Bedos³William Bastian¹Sydney Honold⁵Pablo Pineyro³Aleksandra Pawlak¹Megan P Corbett¹Eugene Douglass¹Karin Allenspach¹Jonathan P Mochel^1*

• ¹University of Georgia, Athens, United States
• ²3D health Solutions, Athens, United States
• ³iowa state university, Athens, United States
• ⁴Moffitt Cancer Center, Tampa, United States
• ⁵The University of Oklahoma Health Sciences, Oklahoma City, United States

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. Therefore, we believe these types of models can expedite drug testing and create a platform for personalized medicine. 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. Furthermore, scRNA-seq was utilized on a subset of the organoids to identify organoid specific transcriptomic signatures including lung, pancreas, kidney, and bladder. 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.