Poultry Production and Product Safety Research Unit, ARS/USDA, Fayetteville, AR, 72701, USA.
Department of Poultry Science, University of Arkansas, Fayetteville, AR, 72701, USA.
BMC Vet Res. 2020 Jun 5;16(1):179. doi: 10.1186/s12917-020-02397-1.
Three-dimensional models of cell culture such as organoids and mini organs accord better advantage over regular cell culture because of their ability to simulate organ functions hence, used for disease modeling, metabolic research, and the development of therapeutics strategies. However, most advances in this area are limited to mammalian species with little progress in others such as poultry where it can be deployed to study problems of agricultural importance. In the course of enterocyte culture in chicken, we observed that intestinal mucosal villus-crypts self-repair and form spheroid-like structures which appear to be useful as ex vivo models to study enteric physiology and diseases.
The villus-crypts harvested from chicken intestinal mucosa were cultured to generate enteroids, purified by filtration then re cultured with different chemicals and growth factors to assess their response based on their morphological dispositions. Histochemical analyses using marker antibodies and probes showed the enteroids consisting different cell types such as epithelial, goblet, and enteroendocrine cells typical to villi and retain functional characteristics of intestinal mucosa.
We present a simple procedure to generate avian crypt-villous enteroids containing different cell types. Because the absorptive cells are functionally positioned outwards, similar to the luminal enterocytes, the cells have better advantages to interact with the factors present in the culture medium. Thus, the enteroids have the potential to study the physiology, metabolism, and pathology of the intestinal villi and can be useful for preliminary screenings of the factors that may affect gut health in a cost-effective manner and reduce the use of live animals.
细胞培养的三维模型，如类器官和微型器官，由于其模拟器官功能的能力优于常规细胞培养，因此更具优势，用于疾病建模、代谢研究和治疗策略的开发。然而，该领域的大多数进展仅限于哺乳动物物种，而在禽类等其他物种中进展甚微，禽类可以用于研究具有农业重要性的问题。在鸡的肠上皮细胞培养过程中，我们观察到肠黏膜绒毛-隐窝会自我修复并形成类似球体的结构，这些结构似乎可用作研究肠生理和疾病的离体模型。
从鸡肠黏膜中采集的绒毛-隐窝被培养以生成肠类器官，然后通过过滤进行纯化，再用不同的化学物质和生长因子重新培养，以根据其形态特征评估它们的反应。使用标记抗体和探针进行组织化学分析表明，肠类器官包含不同的细胞类型，如上皮细胞、杯状细胞和肠内分泌细胞，这些细胞是典型的绒毛细胞，并保留了肠黏膜的功能特征。
我们提出了一种简单的方法来生成含有不同细胞类型的禽类隐窝-绒毛肠类器官。由于吸收细胞在功能上向外排列，类似于腔肠细胞，因此这些细胞与培养基中存在的因子相互作用的优势更好。因此，肠类器官有可能研究肠绒毛的生理学、代谢和病理学，并且可以用于以经济有效的方式初步筛选可能影响肠道健康的因素，减少对活体动物的使用。
