Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays
- Kelli L VanDussen1,
- Jeffrey M Marinshaw2,
- Nurmohammad Shaikh3,
- Hiroyuki Miyoshi1,
- Clara Moon1,
- Phillip I Tarr3,4,
- Matthew A Ciorba2,
- Thaddeus S Stappenbeck1
- 1Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, USA
- 2Division of Gastroenterology, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, USA
- 3Division of Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA
- 4Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, USA
- Correspondence to Dr Thaddeus S Stappenbeck, Department of Pathology and Immunology, Washington University School of Medicine, 660 S. Euclid, Box 8118, St. Louis, MO 63110, USA; and Dr Matthew A Ciorba, Division of Gastroenterology, Department of Internal Medicine, Washington University School of Medicine, 660 S. Euclid, Box 8124, St. Louis, MO 63110, USA; firstname.lastname@example.org
- Received 19 December 2013
- Revised 22 May 2014
- Accepted 6 June 2014
- Published Online First 9 July 2014
Objective 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).
Design 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.
Results 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 epithelial spheroids showed region-specific development of mature epithelial lineages. These cells formed functional, polarised monolayers covered by a secreted mucus layer when grown on Transwell membranes. Using two-dimensional culture, these cells also demonstrated novel adherence phenotypes with various strains of pathogenic Escherichia coli.
Conclusions This culture system will facilitate the study of interindividual, functional studies of human intestinal epithelial cells, including host–microbial interactions.