Establishment of human colonic epithelial cells in long-term culture

Clifford W. Deveney, Leslie Rand-Luby, Michael J. Rutten, Cheryl A. Luttropp, Wendy M. Fowler, John Land, Camie L. Meichsner, Mehrdad Farahmand, Brett C. Sheppard, Richard A. Crass, Karen E. Deveney

Research output: Contribution to journalArticle

39 Scopus citations

Abstract

Study of normal colonic function is important in understanding the cellular mechanisms of carcinogenesis and other diseases of the colon. However, colonic pathophysiological studies have been limited due to the lack of long-term cultures of normal human colonic epithelial cells. The purpose of the present study was to develop methods of isolating viable human colonic epithelial cells for the establishment of nontransformed colonic epithelial cell lines. Human colonic epithelial cells were isolated from surgically resected normal human colons. We found that the use of a short enzymatic digestion gave a consistently higher number (>90%) of viable human colonic epithelial cells. These isolated colonocytes were grown on plastic, collagen- coated filters, or feeder layers using different media formulations. Those colonocytes from the initial primary cultures that were most 'epithelial' in appearance were cloned and passaged to establish long-term cultures of nontransformed human colonic epithelial cells. The epithelial nature and secretory function of these established cell lines were confirmed by morphological criteria (light microscopy, phase contrast microscopy, and electron microscopy). We found that the long-term cultures remained immunopositive to anti-cytokeratin antibodies and immunonegative to anti- vimentin antibodies. Using a soft agar assay we found that the colonocytes did not form colonies, suggesting that the long-term culturing did not cause these cells to become transformed. Under serum-free conditions, we found that epidermal growth factor and transforming growth factor-α were equally potent in their mitogenic effects for these colonocytes. Some of the subcultured cells could be maintained for at least 8 months and still retain their epithelial characteristics. We believe that this methodology will serve as a valuable tool for the isolation and culturing of human colonic epithelial cells for studies of normal and malignant colonic disease processes.

Original languageEnglish (US)
Pages (from-to)161-169
Number of pages9
JournalJournal of Surgical Research
Volume64
Issue number2
DOIs
StatePublished - Aug 1996
Externally publishedYes

ASJC Scopus subject areas

  • Surgery

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    Deveney, C. W., Rand-Luby, L., Rutten, M. J., Luttropp, C. A., Fowler, W. M., Land, J., Meichsner, C. L., Farahmand, M., Sheppard, B. C., Crass, R. A., & Deveney, K. E. (1996). Establishment of human colonic epithelial cells in long-term culture. Journal of Surgical Research, 64(2), 161-169. https://doi.org/10.1006/jsre.1996.0323