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Cited by 525Digestive system and related healthCancer Cells and MetastasisWnt/β-catenin signaling in development and cancer

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

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VanDussen, Kelli L, Marinshaw, Jeffrey M, Shaikh, Nurmohammad, Miyoshi, Hiroyuki, Moon, Clara, Tarr, Phillip I, Ciorba, Matthew A, Stappenbeck, Thaddeus S·2015·Gut
DOI 10.1136/gutjnl-2013-306651PMC PMC4305344
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Abstract

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.

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Cited by

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Authors
VanDussen, K. L., Marinshaw, J. M., Shaikh, N., Miyoshi, H., Moon, C., Tarr, P. I., Ciorba, M. A., & Stappenbeck, T. S.
Affiliations
Washington University in St. Louis (US)
Corresponding author
Stappenbeck, Thaddeus S
ORCID
Venue
Gut
Publisher
BMJ
Published
2014-07-09
Volume
64
Issue
6
Pages
911-920
ISSN
  • 0017-5749 (ISSN-L)
  • 1468-3288
DOI
10.1136/gutjnl-2013-306651
PMC
PMC4305344
OpenAlex
W2147832388
Cited by
525
References
2
Citations by year
2026: 29 · 2025: 45 · 2024: 53 · 2023: 37 · 2022: 57 · 2021: 64 · 2020: 60 · 2019: 53 · 2018: 50 · 2017: 37 · 2016: 23 · 2015: 14 · 2014: 2
FWCI
16.64
Type
journal article
Language
en
Open access
Locations
  • Gut (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • Crohn's and Colitis Foundation of America
  • Crohn's and Colitis Foundation
  • National Institutes of Health
  • National Institute of Allergy and Infectious Diseases (U01AI095550, AI095550, T32AI007163)
  • National Institute of Diabetes and Digestive and Kidney Diseases (R03DK100737, DK100737, DK90251, K08DK089016, R01DK071619, DK089016, U54DK102557, DK102557, DK071619, P30DK052574)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Digestive system and related health
  • Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
Keywords
human intestinal epithelial cells, patient-derived cell lines, intestinal stem cells, Wnt3a, R-spondin, noggin, Notch signaling inhibition, epithelial differentiation, spheroids, mucus secretion, host-microbe interactions, individualised medicine, endoscopic biopsy
MeSH terms
Analysis of Variance, Animals, Bacterial Adhesion, Biological Assay, Biopsy, Cell Differentiation, Cell Line, Cells, Cultured, Epithelial Cells, Genetic Markers, Humans, Ileum, Intestinal Mucosa, Rectum, RNA, Messenger, Reproducibility of Results, Culture Media, Conditioned, Spheroids, Cellular, Cell Culture Techniques, Mice

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