Cell Biology and Toxicology

, Volume 21, Issue 1, pp 1–26

The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics


  • Y. Sambuy
    • Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione (INRAN)CNR
  • I. De Angelis
    • Istituto Superiore di SanitàCNR
  • G. Ranaldi
    • Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione (INRAN)CNR
  • M. L. Scarino
    • Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione (INRAN)CNR
  • A. Stammati
    • Istituto Superiore di SanitàCNR
    • Istituto di Neurobiologia e Medicina MolecolareCNR

DOI: 10.1007/s10565-005-0085-6

Cite this article as:
Sambuy, Y., De Angelis, I., Ranaldi, G. et al. Cell Biol Toxicol (2005) 21: 1. doi:10.1007/s10565-005-0085-6


The human intestinal Caco-2 cell line has been extensively used over the last twenty years as a model of the intestinal barrier. The parental cell line, originally obtained from a human colon adenocarcinoma, undergoes in culture a process of spontaneous differentiation that leads to the formation of a monolayer of cells, expressing several morphological and functional characteristics of the mature enterocyte. Culture-related conditions were shown to influence the expression of these characteristics, in part due to the intrinsic heterogeneity of the parental cell line, leading to selection of sub-populations of cells becoming prominent in the culture. In addition, several clonal cell lines have been isolated from the parental line, exhibiting in general a more homogeneous expression of differentiation traits, while not always expressing all characteristics of the parental line. Culture-related conditions, as well as the different Caco-2 cell lines utilized in different laboratories, often make it extremely difficult to compare results in the literature. This review is aimed at summarizing recent, or previously unreviewed, data from the literature on the effects of culture-related factors and the influence of line sub-types (parental vs. different clonal lines) on the expression of differentiation traits important for the use of Caco-2 cells as a model of the absorptive and defensive properties of the intestinal mucosa. Since the use of Caco-2 cells has grown exponentially in recent years, it is particularly important to highlight these methodological aspects in order to promote the standardization and optimisation of this intestinal model.


biotransformation activitiesCaco-2clonal cell linesculture conditionsdifferentiationtransporters

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© Springer Science + Business Media, Inc. 2005