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Chronic Exposure To Microbial Stimuli Affects The Development Of The Intestinal Epithelium In Human Colonic Enteroids
Author(s) -
Rodrigues Ely,
Slobbe Lynn,
Schultz Michael,
Butt Grant
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.999.7
Subject(s) - lgr5 , goblet cell , biology , stem cell , microbiology and biotechnology , intestinal epithelium , epithelium , western blot , blot , cancer stem cell , gene , biochemistry , genetics
Adult stem cells and transient amplifying cells that support epithelial regeneration are key sites where bacteria could modulate epithelial regeneration and homeostasis. The aim of this research was to determine if modulation of adult intestinal stem cells by the bacterial components affected the development of the intestinal epithelium. Enteroids were grown from crypts isolated from the transverse colon of healthy individuals and transferred to Matrigel and growth media for 15 days in the presence and absence of lipopolysaccharide (LPS, 20 ng ml ‐1 ). Organoid structure was assessed by light microscopy and gene expression by microarray, qPCR, Western Blotting and immunohistochemistry (IHC). LPS increased the total number of goblet cells to 20% + 3% (n=5), compared with none in the untreated controls. Microarray analysis showed an increase in the goblet cell associated proteins MUC2, Clca1, Zg16, Mpgc60, microtubule‐associated protein and resistin‐like beta protein and also an increase in genes associated with cell cycle, growth and differentiation in the LPS treated enteroids. Western blot confirmed an increase in the goblet cell marker MUC2. IHC confirmed an increase in the number of MUC2 and TFF3 positive cells confirming a lineage change to give more goblet cells. Collectively, these data indicate that LPS modulates stem cell activity and effects intestinal epithelial development.