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

Cell Biol Toxicol. 2005 Jan;21(1):1-26. 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Autocrine Communication / physiology
  • Caco-2 Cells
  • Cell Adhesion / drug effects
  • Cell Adhesion / physiology
  • Cell Count
  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology
  • Cell Physiological Phenomena*
  • Clone Cells / cytology
  • Clone Cells / drug effects
  • Clone Cells / physiology
  • Culture Media / chemistry
  • Culture Media / pharmacology
  • Enterocytes / cytology
  • Enterocytes / drug effects
  • Enterocytes / physiology
  • Extracellular Matrix / metabolism
  • Humans
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / drug effects
  • Intestinal Mucosa / physiology*
  • Membrane Transport Proteins / metabolism
  • Paracrine Communication / physiology
  • Sucrase-Isomaltase Complex / metabolism


  • Culture Media
  • Membrane Transport Proteins
  • Sucrase-Isomaltase Complex