Organotypical tissue cultures from adult murine colon as an in vitro model of intestinal mucosa.
Together with animal experiments, organotypical cell cultures are important models for analyzing cellular interactions of the mucosal epithelium and pathogenic mechanisms in the gastrointestinal tract. Here, we introduce a three-dimensional culture model from the adult mouse colon for cell biological investigations in an in vivo-like environment. These explant cultures were cultured for up to 2 weeks and maintained typical characteristics of the intestinal mucosa, including a high-prismatic epithelium with specific epithelial cell-to-cell connections, a basal lamina and various connective tissue cell types, as analyzed with immunohistological and electron microscopic methods. The function of the epithelium was tested by treating the cultures with dexamethasone, which resulted in a strong upregulation of the serum- and glucocorticoid-inducible kinase 1 similar to that found in vivo. The culture system was investigated in infection experiments with the fungal pathogen Candida albicans. Wildtype but not Deltacph1/Deltaefg1-knockout Candida adhered to, penetrated and infiltrated the epithelial barrier. The results demonstrate the potential usefulness of this intestinal in vitro model for studying epithelial cell-cell interactions, cellular signaling and microbiological infections in a three-dimensional cell arrangement.
Bareiss
Petra M
PM
Institute of Anatomy, Center for Regenerative Biology and Medicine, Eberhardt-Karls-University Tuebingen, Oesterbergstrasse 3, 72074, Tuebingen, Germany.
Metzger
Marco
M
Sohn
Kai
K
Rupp
Steffen
S
Frick
Julia S
JS
Autenrieth
Ingo B
IB
Lang
Florian
F
Schwarz
Heinz
H
Skutella
Thomas
T
Just
Lothar
L
D000818:Animals; D002176:Candida albicans / Q000235:genetics / Q000378:metabolism; D002177:Candidiasis / Q000378:metabolism / Q000382:microbiology; D018929:Cell Culture Techniques; D003106:Colon / Q000187:drug effects / Q000382:microbiology / Q000648:ultrastructure*; D003907:Dexamethasone / Q000494:pharmacology; D004848:Epithelium / Q000187:drug effects / Q000382:microbiology / Q000648:ultrastructure; D017874:Immediate-Early Proteins / Q000378:metabolism; D007413:Intestinal Mucosa / Q000187:drug effects / Q000382:microbiology / Q000648:ultrastructure*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D046509:Tissue Culture Techniques; D000099184:Serum-Glucocorticoid Regulated Kinases
D003907:Dexamethasone; D017874:Immediate-Early Proteins; D017346:Protein Serine-Threonine Kinases; D000099184:Serum-Glucocorticoid Regulated Kinases