不同区段小鼠小肠类器官体外培养方法建立

Establishment of ex Vivo Culture Method of Small Intestinal Organoids of Different Segments of Mouse

肠道是食物营养物质消化吸收的主要器官,良好的肠道体外培养模型有助于推动新型食物营养物质及功能活性物质的吸收及功效评价方法建立。本文以含有肠道干细胞的肠道隐窝为基础,通过优化不同区段小肠隐窝分离方法和体外培养基组成,建立一套完整的不同区段小肠类器官(包括十二指肠、空肠和回肠)培养方法。采用免疫荧光和RT-PCR方法进一步证明不同区段小肠类器官与相应肠道组织具有相似结构特征及细胞组成,而且这些性状在传代过程中具有稳定性。结论:本文建立的不同区段小肠类器官培养方法,可能为食品领域研究营养物质功能评价及吸收评价提供良好的体外研究工具。

Intestinal tract is the major organ for digestion and absorption of food nutrients.The establishment of an effective intestinal ex vivo model is helpful to promote the development of a novel evaluation system for absorption and efficacy of food nutrients and functional active substances.Based on the intestinal crypt containing intestinal stem cells,we created an integrated set of organoid culture of different intestinal segments(including duodenum,jejunum and ileum)of mouse through optimizing the isolation method of intestinal crypt from different intestinal segment and the composition of culture medium.In addition,RT-PCR and immunofluorescence results demonstrated that the organoids derived from different segment recapitulate the structural characteristics and cell composition of corresponding intestinal tissues.These characters had high stability during in vitro repeated passage.In short,the establishment of orgnaoid culture of different intestinal segment may provide a promising ex vivo model for the evaluation of absorption and function of food nutrients in the food research field.