Immortalized epithelial cells derived from human colon biopsies express stem cell markers and differentiate in vitro

Gastroenterology. 2010 Mar;138(3):1012-21.e1-5. doi: 10.1053/j.gastro.2009.11.052. Epub 2009 Dec 4.


Background & aims: Long-term propagation of human colonic epithelial cells (HCEC) of adult origin has been a challenge; currently used HCEC lines are of malignant origin and/or contain multiple cytogenetic changes. We sought to immortalize human colon biopsy-derived cells expressing stem cell markers and retaining multilineage epithelial differentiation capability.

Methods: We isolated and cultured cells from biopsy samples of 2 patients undergoing routine screening colonoscopy. Cells were immortalized by expression of the nononcogenic proteins cyclin-dependent kinase 4 (Cdk4) and the catalytic component of human telomerase (hTERT) and maintained for more than 1 year in culture.

Results: The actively proliferating HCECs expressed the mesenchymal markers vimentin and alpha-smooth muscle actin. Upon growth arrest, cells assumed a cuboidal shape, decreased their mesenchymal features, and expressed markers of colonic epithelial cells such as cytokeratin 18, zonula occludens-1, mucins-1 and -2, antigen A33, and dipeptidyl peptidase 4. Immortalized cells expressed stem cell markers that included LGR5, BMI1, CD29, and CD44. When placed in Matrigel in the absence of a mesenchymal feeder layer, individual cells divided and formed self-organizing, cyst-like structures; a subset of cells exhibited mucin-2 or polarized villin staining.

Conclusions: We established immortalized HCECs that are capable of self-renewal and multilineage differentiation. These cells should serve as valuable reagents for studying colon stem cell biology, differentiation, and pathogenesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Biomarkers / metabolism*
  • Biopsy
  • Cell Differentiation*
  • Cell Line
  • Cell Lineage*
  • Cell Proliferation
  • Cell Shape
  • Collagen / metabolism
  • Colon / metabolism*
  • Colon / ultrastructure
  • Colonoscopy
  • Cyclin-Dependent Kinase 4 / genetics
  • Drug Combinations
  • Epithelial Cells / metabolism*
  • Epithelial Cells / ultrastructure
  • Humans
  • Karyotyping
  • Laminin / metabolism
  • Proteoglycans / metabolism
  • Stem Cells / metabolism*
  • Stem Cells / ultrastructure
  • Telomerase / genetics
  • Time Factors
  • Transfection


  • Biomarkers
  • Drug Combinations
  • Laminin
  • Proteoglycans
  • matrigel
  • Collagen
  • CDK4 protein, human
  • Cyclin-Dependent Kinase 4
  • TERT protein, human
  • Telomerase