Immortalized colonic epithelial progenitor cells express stem cell markers and differentiate in vitro

ABSTRACT: Immortalized colonic epithelial progenitor cells derived from normal human colon biopsies express stem cell markers and differentiate in vitro The sustained growth of normal non-transformed human colonic epithelial cells has historically been exceptionally challenging. We here report the successful long-term propagation of colon biopsy derived cells expressing both epithelial and mesenchymal features, intestinal stem cell markers, and demonstrating multilineage epithelial differentiation capability. Cells isolated from the biopsies of two patients undergoing routine screening colonoscopy have been successfully passaged in vitro, subsequently immortalized with non-oncogenic proteins (cyclin dependent kinase 4 [CDK4] and the catalytic component of human telomerase [hTERT]), and maintained for well over a year in continuous cell culture. Immunofluorescence experiments reveal the immortalized populations of cells significantly express the progenitor epithelial cell marker mucin-1 and the colon epithelial specific marker A33. In addition, staining shows subsets of cells express chromogranin A, mucin-2, and dipeptyl peptidase 4, corresponding to neuroendocrine, mucus secreting, and absorptive colonic cell lineages, respectively. The cells also possess some mesenchymal features, including vimentin and α- smooth muscle actin expression when proliferating actively. Finally, immortalized cells express various stem cell markers including LGR5, BMI1, CD29, and CD44. When cells are seeded at low density in Matrigel® in the absence of a mesenchymal feeder layer individual cells are able to divide and form self-organizing, cyst-like structures with a subset of cells exhibiting either mucin-2 or polarized villin staining. These findings demonstrate that the immortalized human colonic cells are capable of self renewal and multilineage differentiation. These cells should serve as valuable cellular reagents for studying normal colon stem cell biology, differentiation, and disease development.

ORGANISM(S): Homo sapiens

PROVIDER: GSE18433 | GEO | 2009/10/07

SECONDARY ACCESSION(S): PRJNA118233

REPOSITORIES: GEO