Immortalized Epithelial Cells Derived From Human Colon Biopsies Express Stem Cell Markers and Differentiate In Vitro

Andres I. Roig, Ugur Eskiocak, Suzie K. Hight, Sang Bum Kim, Oliver Delgado, Rhonda F. Souza, Stuart J. Spechler, Woodring E. Wright, Jerry W. Shay

Research output: Contribution to journalArticlepeer-review

135 Scopus citations


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 α-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.

Original languageEnglish (US)
Pages (from-to)1012-1021.e5
Issue number3
StatePublished - Mar 2010


  • Epithelial Mesenchymal Transition
  • Multilineage Differentiation
  • Self-Renewal
  • Stem/Progenitor Cells

ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology


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