Characterization of discrete equine intestinal epithelial cell lineages

Abstract

OBJECTIVE To characterize epithelial cells of the small intestine and colon in horses without clinical gastrointestinal abnormalities with an emphasis on the stem cell niche constituents. SAMPLE Mucosal biopsy specimens from small and large intestines obtained from 12 horses euthanized for reasons unrelated to gastrointestinal disease or systemic disease. PROCEDURES Intestinal biopsy specimens were collected by sharp dissection immediately following euthanasia. Specimens were prepared for immunohistochemical, immunofluorescence, and transmission electron microscopic imaging to detect and characterize each epithelial cell type. Antibodies against protein biomarkers for cellular identification were selected on the basis of expression in other mammalian species. RESULTS Intestinal epithelial cell types were identified by means of immunostaining and morphological characterization with transmission electron microscopy. Some differences in biomarker expression and antibody cross-reactivity were identified in equine tissue, compared with other species. However, each known type of mucosal epithelial cell was identified in equine tissue. CONCLUSIONS AND CLINICAL RELEVANCE The methodology used can enhance detection of stem cells and progenitor cells as well as postmitotic cell lineages in equine intestinal tissues. Results may have relevance to regenerative potential of intestinal mucosa and survival in horses with colic.

Advanced three‐dimensional culture of equine intestinal epithelial stem cells

“…Several approaches to identify equine epithelial cell types were pursued because of the innate advantages and disadvantages of each technique. Similar to other studies, antibody-based assays alone failed to positively identify all intestinal epithelial cell types [9,16]. The antibodies that were used were commercially derived and raised against proteins in species other than horses.…”

“…Many cellular biomarkers were conserved between species, as indicated by cross-reactivity of several antibodies with equine proteins in this study. A previous study helped to establish the existence and normal distribution of cell types within the equine small and large intestinal mucosa and the reagents and tools currently available [16]. Successful amplification of known gene cellular biomarkers was further used to characterise and confirm the existence of all known cell types that exist in the equine intestinal epithelium.…”

“…Enteroids were permeabilised in a 0.3% Triton X-100 PBS solution for 20 min and then blocked in protein block solution k for 30 min. Primary antibodies labeling individual cell types were applied at concentrations previously published [16], and incubated overnight at 4°C. Additional primary antibodies included anti-Lysozyme and anti-Ki67 for Paneth cells and proliferating cell populations, respectively.…”

Algal Biofuels: Current Status and Key Challenges

“…Biorefining means breaking down biomass into bioproducts-either fuel or nonfuel products-via sustainable methods [181]. Depending on the target product, a variety of refinery schemes are available [182].…”

Non-invasive evaluation of the equine gastrointestinal mucosal transcriptome

“…cell types present in these data, we reviewed the literature for marker genes expressed either solely by a specific cell type or at least highly enriched in a specific cell type [19][20][21][22][23][24][25][26][27][28][29][30][31][32]. In particular, we extracted the numbers of reads in each sample for the following cell types: intestinal stem cells, absorptive cells, transit amplifying cells, Paneth cells, tuft cells, goblet cells, macrophages, lymphocytes, neutrophils, and smooth muscle cells.…”