Production and characterization of avian crypt-villus enteroids and the effect of chemicals
Production and characterization of avian crypt-villus enteroids and the effect of chemicals
复制标题
DOI：
10.1186/s12917-020-02397-1
复制
发表时间：
2020-06-05
影响因子：
2.6
通讯作者：
Rath, Narayan C.
中科院分区：
文献类型：
--
作者：
Acharya, Mohan;Arsi, Komala;Rath, Narayan C.
背景三维细胞培养模型，如类器官和微型器官，由于其能够模拟器官功能而比常规细胞培养具有更好的优势，因此被用于疾病建模、代谢研究和治疗策略的开发。然而，这一领域的大多数进展仅限于哺乳动物物种，而在家禽等其他物种方面进展甚微，可用于研究具有农业重要性的问题。在鸡肠道细胞培养过程中，我们观察到肠粘膜绒毛-隐窝具有自我修复和形成类球形结构的能力，可作为研究肠道生理和疾病的体外模型。结果从鸡肠黏膜获取的绒毛-隐窝培养生成肠样物质，过滤纯化后再用不同的化学物质和生长因子重新培养，根据其形态特征评估其反应。标记抗体和探针的组织化学分析显示，小肠样体由不同的细胞类型组成，如绒毛典型的上皮细胞、杯状细胞和肠内分泌细胞，并保留了肠粘膜的功能特征。由于吸收细胞在功能上是向外定位的，类似于腔内肠细胞，因此细胞在与培养液中存在的因子相互作用方面具有更好的优势。因此，肠样体有可能研究肠道绒毛的生理、代谢和病理，并可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，并减少活体动物的使用。
BackgroundThree-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.ResultsThe 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.ConclusionsWe 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.