Immortalization of adult human colonic epithelial cells extracted from normal tissues obtained via colonoscopy
Protocol Exchange. 2010 February 16;
- Abstract
- Many of the models currently available to study colon cell biology and colonic diseases are based on in vivo approaches (mouse models) or use colonic epithelial cells that can be easily maintained in culture, that is, cells of a cancerous origin1. It is necessary to develop long-term culture models of non-transformed human colonic epithelial cells (HCEC) to further enhance our understanding of important human colonic diseases, such as colorectal cancer (CRC) progression, that would be difficult to achieve with the current systems available for study. In this protocol, we present the methodology of a recently developed procedure for experimentally immortalizing adult human colonic epithelial progenitor cells for long-term growth2. The described protocol contains aspects of methods previously established for the short-term culture of mammalian intestinal cells as well as other techniques used to enhance cellular viability, proliferative potential, and promote long-term growth of other epithelial cells without de novo creation of oncogenic mutations or alterations of cellular karyotype3-7. The cells we have immortalized with this protocol have been replicating for well over a year, have colonic epithelial progenitor cell features, retain a diploid karyotype and do not exhibit tumorigenic features, and tolerate repeated freezing and thawing.
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