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Colonic Crypt‐Derived 3D Gut Organoid from Hypertensive Rat Maintain In vivo Characteristics
Author(s) -
li Jing,
Su Qing,
Karas Marianthi M.,
Richards Elaine M.,
Raizada Mohan K.
Publication year - 2020
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.2020.34.s1.09495
Subject(s) - organoid , crypt , in vivo , epithelium , gut flora , biology , medicine , matrigel , endocrinology , pathology , microbiology and biotechnology , immunology
Objective Substantial evidence links gut dysbiosis to hypertension in both animal models and in patients with high blood pressure. Increased blood pressure and hypertension‐associated pathologies in normotensive animals transplanted with fecal microbiota from hypertensive animals, support the concept of microbiota‐gut communication as an important early signal initiating and establishing hypertension. We hypothesized that colonic cryp‐3D organoids would retain the characteristics of the in vivo gut epithelium form which they were derived, and would provide a model system to investigate epithelial‐microbiota communication in hypertension. Methods Colonic crypts were isolated from adult Wistar‐Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). 3D organoid cultures were established in organoid growth medium with Matrigel matrix and cultured for 3, 5 and 7 days without passage. The shapes and number of organoids were assessed by bright‐field microscopy using Trypan blue. Growth, differentiation and cell types in organoids were characterized with the use of antibodies (Ki67, Alpi, KRT20 and Muc2). Data were compared between WKY rats and SHR organoids. Results Reproducible 3D organoid cultures were successfully established from the crypts of adult WKY and SHR. They proliferated and differentiated, and predominately contained enterocytes and goblet cells. We observed significantly more organoids per crypt from normotensive vs hypertensive rats (WKY: 0.20 vs. SHR: 0.11; p<0.01). In vivo , WKY rats had 18% longer colons than SHR (0.07 cm/g vs. 0.059 cm/g; p<0.0001). Conclusions These observations demonstrate that 3D organoid cultures maintain characteristics of the colonic epithelium and show significant differences between WKY rats and SHR. Thus, these organoids represent an ideal ex vivo system to investigate the gut epithelium in hypertension, and to examine the effects of gut microbiome‐derived metabolites in blood pressure control and initiation and establishment of hypertension. Support or Funding Information HL132448