Jul 11, 2014

Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays

Gut
K L VanDussenThaddeus S Stappenbeck

Abstract

The technology for the growth of human intestinal epithelial cells is rapidly progressing. An exciting possibility is that this system could serve as a platform for individualised medicine and research. However, to achieve this goal, human epithelial culture must be enhanced so that biopsies from individuals can be used to reproducibly generate cell lines in a short time frame so that multiple, functional assays can be performed (ie, barrier function and host-microbial interactions). We created a large panel of human gastrointestinal epithelial cell lines (n=65) from patient biopsies taken during routine upper and lower endoscopy procedures. Proliferative stem/progenitor cells were rapidly expanded using a high concentration of conditioned media containing the factors critical for growth (Wnt3a, R-spondin and Noggin). A combination of lower conditioned media concentration and Notch inhibition was used to differentiate these cells for additional assays. We obtained epithelial lines from all accessible tissue sites within 2 weeks of culture. The intestinal cell lines were enriched for stem cell markers and rapidly grew as spheroids that required passage at 1:3-1:4 every 3 days. Under differentiation conditions, intestinal epithel...Continue Reading

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Citations

Mentioned in this Paper

NOG gene
Endoscopy (Procedure)
Tissue Membrane
RSPO1 gene
Notch
Pathogenic Organism
Culture Media, Conditioned
Squamous Transitional Epithelial Cell Count
WNT3A gene
Ileum

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