title: Immortalized epithelial cells derived from human colon biopsies express stem cell markers and differentiate in vitro.
abstract: OBJECTIVE
  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.
authors: Department of Cell Biology, Division of Digestive and Liver Diseases, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
authors: Andres I
authors: AI
authors: Roig
authors: Ugur
authors: U
authors: Eskiocak
authors: Suzie K
authors: SK
authors: Hight
authors: Sang Bum
authors: SB
authors: Kim
authors: Oliver
authors: O
authors: Delgado
authors: Rhonda F
authors: RF
authors: Souza
authors: Stuart J
authors: SJ
authors: Spechler
authors: Woodring E
authors: WE
authors: Wright
authors: Jerry W
authors: JW
authors: Shay
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D001706:Biopsy; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D049109:Cell Proliferation; D048430:Cell Shape; D003094:Collagen / Q000378:metabolism; D003106:Colon / Q000378:metabolism* / Q000648:ultrastructure; D003113:Colonoscopy; D051358:Cyclin-Dependent Kinase 4 / Q000235:genetics; D004338:Drug Combinations; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007621:Karyotyping; D007797:Laminin / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism* / Q000648:ultrastructure; D019098:Telomerase / Q000235:genetics; D013997:Time Factors; D014162:Transfection
chemical_list: D015415:Biomarkers; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4; C509186:TERT protein, human; D019098:Telomerase