For decades, two-dimensional (2D) cell cultures have been used as the gold standard in vitro models for basic research, drug discovery and preclinical screening applications. However, since 2009, the field has quickly shifted towards the use of three-dimensional (3D) stem cell-derived organoids. Organoids offer a variety of advantages compared with conventional 2D models, including being self-organized, retaining their cellular polarity, and being simplified versions of organs in vitro. Adult stem cells can self-renew, differentiate into multiple cell types, and are genomically stable over multiple passage lines, providing a variety of technical benefits and offering the closest recapitulation of the in vivo environment. Our laboratory has focused on expanding, characterizing, and utilizing animal-derived 3D organoid models, specifically focusing on canines; but also including other animals such as swine, turtles, snakes, and squirrels to study their unique adaptations. Practically, we can use 3D canine colonoids for the screening of anti-inflammatory candidates using our tumor necrosis factor (TNF)-α stimulation model. Additionally, we have grown and used swine lung organoids as a model for studying H1N1 viral uptake. Furthermore, we have grown and bio-banked tumor organoids/spheroids from a variety of canine cancers, including urothelial carcinoma (derived from urine and biopsies) and nasal carcinoma, among others, with future applications in precision veterinary medicine and comparative oncology. To accomplish this goal, we have carried out in vitro screening of a variety of chemotherapies and combination therapies at different concentrations across patients to predict in vivo responses and demonstrate the predictive ability of the organoid model. Finally, our laboratory has derived and characterized a variety of healthy organoid lines from canines including the pancreas, lung, liver, bladder, colon, and pituitary to act as healthy controls in mechanistic studies looking at disease pathogenesis and for uses in basic biology. In summary, in vitro organoid models from various animal species and tissues can be used in a variety of basic and applied research fields, including virology, toxicology, and oncology, with the potential to broaden our scientific understanding of disease and overall improve human and veterinary medicine.

中文翻译：

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166 动物研究中非模型类器官的开发：体外模型的范式转变


几十年来，二维 （2D） 细胞培养一直被用作基础研究、药物发现和临床前筛选应用的体外模型金标准。然而，自 2009 年以来，该领域迅速转向使用三维 （3D） 干细胞衍生的类器官。与传统的 2D 模型相比，类器官具有多种优势，包括自组织、保留其细胞极性以及在体外是器官的简化版本。成体干细胞可以自我更新，分化为多种细胞类型，并且在多条传代系上保持基因组稳定性，提供多种技术优势，并提供最接近体内环境的概括。我们的实验室专注于扩展、表征和利用动物来源的 3D 类器官模型，特别关注犬科动物;但也包括猪、、蛇和松鼠等其他动物，以研究它们独特的适应能力。实际上，我们可以使用 3D 犬结肠来使用我们的肿瘤坏死因子 （TNF） α刺激模型来筛选抗炎候选药物。此外，我们还培养并使用猪肺类器官作为研究 H1N1 病毒摄取的模型。此外，我们已经从各种犬癌中培养和生物储存了肿瘤类器官/球状体，包括尿路上皮癌（来源于尿液和活检）和鼻癌等，未来将在精准兽医学和比较肿瘤学中应用。为了实现这一目标，我们对患者体内不同浓度的各种化疗和联合疗法进行了体外筛选，以预测体内反应并证明类器官模型的预测能力。 最后，我们的实验室从犬科动物（包括胰腺、肺、肝脏、膀胱、结肠和垂体）中衍生并表征了各种健康的类器官系，以作为研究疾病发病机制的机制研究和基础生物学的用途的健康对照。总之，来自各种动物物种和组织的体外类器官模型可用于各种基础和应用研究领域，包括病毒学、毒理学和肿瘤学，有可能拓宽我们对疾病的科学理解，并全面改善人类和兽医学。