Publications

@article{villegas-novoa_development_2022,

title = {Development of a Primary Human Intestinal Epithelium Enriched in L-Cells for Assay of GLP-1 Secretion},

author = {Cecilia Villegas-Novoa and Yuli Wang and Christopher E. Sims and Nancy L. Allbritton},

doi = {10.1021/acs.analchem.2c00912},

issn = {1520-6882},

year  = {2022},

date = {2022-07-12},

urldate = {2022-07-12},

journal = {Development},

volume = {94},

number = {27},

pages = {9648–9655},

abstract = {Type 2 diabetes mellitus is a chronic disease associated with obesity and dysregulated human feeding behavior. The hormone glucagon-like peptide 1 (GLP-1), a critical regulator of body weight, food intake, and blood glucose levels, is secreted by enteroendocrine L-cells. The paucity of L-cells in primary intestinal cell cultures including organoids and monolayers has made assays of GLP-1 secretion from primary human cells challenging. In the current paper, an analytical assay pipeline consisting of an optimized human intestinal tissue construct enriched in L-cells paired with standard antibody-based GLP-1 assays was developed to screen compounds for the development of pharmaceuticals to modulate L-cell signaling. The addition of the serotonin receptor agonist Bimu 8, optimization of R-spondin and Noggin concentrations, and utilization of vasoactive intestinal peptide (VIP) increased the density of L-cells in a primary human colonic epithelial monolayer. Additionally, the incorporation of an air-liquid interface culture format increased the L-cell number so that the signal-to-noise ratio of conventional enzyme-linked immunoassays could be used to monitor GLP-1 secretion in compound screens. To demonstrate the utility of the optimized analytical method, 21 types of beverage sweeteners were screened for their ability to stimulate GLP-1 secretion. Stevioside and cyclamate were found to be the most potent inducers of GLP-1 secretion. This platform enables the quantification of GLP-1 secretion from human primary L-cells and will have broad application in understanding L-cell formation and physiology and will improve the identification of modulators of human feeding behavior.},

keywords = {Diabetes Mellitus, Enteroendocrine Cells, Glucagon-Like Peptide 1, Intestinal Mucosa, L Cells},

pubstate = {published},

tppubtype = {article}

}

@article{wang_enterochromaffin_2020,

title = {Enterochromaffin Cell-Enriched Monolayer Platform for Assaying Serotonin Release from Human Primary Intestinal Cells},

author = {Yuli Wang and Christopher E. Sims and Nancy L. Allbritton},

doi = {10.1021/acs.analchem.0c02016},

issn = {1520-6882},

year  = {2020},

date = {2020-09-15},

volume = {92},

number = {18},

pages = {12330–12337},

abstract = {Enteroendocrine (EE) cells within the intestinal epithelium produce a range of hormones that have key roles in modulating satiety and feeding behavior in humans. The regulation of hormone release from EE cells as a potential therapeutic strategy to treat metabolic disorders is highly sought after by the pharmaceutical industry. However, functional studies are limited by the scarcity of EE cells (or surrogates) in both in vivo and in vitro systems. Enterochromaffin (EC) cells are a subtype of EE cells that produce serotonin (5HT). Here, we explored simple strategies to enrich EC cells in in vitro monolayer systems derived from human primary intestinal stem cells. During differentiation of the monolayers, the EC cell lineage was significantly altered by both the culture method [air-liquid interface (ALI) vs submerged] and the presence of vasoactive intestinal peptide (VIP). Compared with traditional submerged cultures without VIP, VIP-assisted ALI culture significantly boosted the number of EC cells and their 5HT secretion by up to 430 and 390%, respectively. The method also increased the numbers of other subtypes of EE cells such as L cells. Additionally, this method generated monolayers with enhanced barrier integrity, so that directional (basal or apical) 5HT secretion was measurable. For all donor tissues, the enriched EC cells improved the signal-to-background ratio and reliability of 5HT release assays. The enhancement in the 5HT secretion behavior was consistent over time from a single donor, but significant variation in the amount of secreted 5HT was present among tissues derived from five different donors. To demonstrate the utility of the EC-enriched monolayer system, 13 types of pungent food ingredients were screened for their ability to stimulate 5HT secretion. Curcumin found in the spice turmeric derived from the Curcuma longa plant was found to be the most potent secretagogue. This EC-enriched cell monolayer platform can provide a valuable analytical tool for the high-throughput screening of nutrients and gut microbial components that alter the secretion of 5HT.},

keywords = {Enterochromaffin Cells, Humans, Intestinal Mucosa, Serotonin},

pubstate = {published},

tppubtype = {article}

}

@article{wang_building_2019,

title = {Building a Thick Mucus Hydrogel Layer to Improve the Physiological Relevance of In Vitro Primary Colonic Epithelial Models},

author = {Y. Wang and R. Kim and C. E. Sims and N. L. Allbritton},

doi = {10.1016/j.jcmgh.2019.07.009},

issn = {2352-345X},

year  = {2019},

date = {2019-01-01},

journal = {Cellular and Molecular Gastroenterology and Hepatology},

volume = {8},

number = {4},

pages = {653–655.e5},

keywords = {Cell Culture Techniques, Colon, Humans, Hydrogels, Intestinal Mucosa, Mucus},

pubstate = {published},

tppubtype = {article}

}