Using primary murine intestinal enteroids to study dietary TAG absorption, lipoprotein synthesis, and the role of apoC-III in the intestine

Javeed Jattan,Cayla Rodia,Diana Li,Adama Diakhate,Hongli Dong,Amy Bataille,Noah F Shroyer,Alison B Kohan

doi:10.1194/jlr.m071340

Javeed Jattan, Cayla Rodia + Show 6 more

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https://doi.org/10.1194/jlr.m071340

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Abstract

Since its initial report in 2009, the intestinal enteroid culture system has been a powerful tool used to study stem cell biology and development in the gastrointestinal tract. However, a major question is whether enteroids retain intestinal function and physiology. There have been significant contributions describing ion transport physiology of human intestinal organoid cultures, as well as physiology of gastric organoids, but critical studies on dietary fat absorption and chylomicron synthesis in primary intestinal enteroids have not been undertaken. Here we report that primary murine enteroid cultures recapitulate in vivo intestinal lipoprotein synthesis and secretion, and reflect key aspects of the physiology of intact intestine in regard to dietary fat absorption. We also show that enteroids can be used to elucidate intestinal mechanisms behind CVD risk factors, including tissue-specific apolipoprotein functions. Using enteroids, we show that intestinal apoC-III overexpression results in the secretion of smaller, less dense chylomicron particles along with reduced triacylglycerol secretion from the intestine. This model significantly expands our ability to test how specific genes or genetic polymorphisms function in dietary fat absorption and the precise intestinal mechanisms that are critical in the etiology of metabolic disease.

Highlights

Since its initial report in 2009, the intestinal enteroid culture system has been a powerful tool used to study stem cell biology and development in the gastrointestinal tract
Whereas Caco-2 cells grow in a 2D monolayer composed solely of epithelial cells, mature enteroids are composed of multiple cell types, including stem cells, goblet cells, and enterocytes [17, 23]
Compared with freshly isolated intestine (Fig. 2A), enteroids significantly decreased markers of terminal epithelial differentiation [brush border villin (VIL1) and enterocyte cytokeratin (KRT)20] from the first day of crypt culture through maturation, though they did not have a significant decrease in enterocyte-specific alkaline phosphatase (ALPI) during this time period

Summary

Introduction

Since its initial report in 2009, the intestinal enteroid culture system has been a powerful tool used to study stem cell biology and development in the gastrointestinal tract. We report that primary murine enteroid cultures recapitulate in vivo intestinal lipoprotein synthesis and secretion, and reflect key aspects of the physiology of intact intestine in regard to dietary fat absorption. We show that intestinal apoC-III overexpression results in the secretion of smaller, less dense chylomicron particles along with reduced triacylglycerol secretion from the intestine This model significantly expands our ability to test how specific genes or genetic polymorphisms function in dietary fat absorption and the precise intestinal mechanisms that are critical in the etiology of metabolic disease.—Jattan, J., C. Despite the importance of the intestine in raising plasma TAG levels in the postprandial state and in apolipoprotein secretion, mechanistic studies are difficult to carry out This is due to a lack of suitable ex vivo cell culture models that are nontransformed yet stable enough in culture for extended studies and genetic manipulations.

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