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Cytotechnology

, Volume 13, Issue 3, pp 175–184 | Cite as

Epithelial properties of human intestinal Caco-2 cells cultured in a serum-free medium

  • Kei Hashimoto
  • Makoto Shimizu
Original Research Papers

Abstract

Human intestinal Caco-2 cells were cultured under serum-free conditions on an insoluble collagen and FCS matrix (Caco-2-SF), and a comparison was made between several characteristics of Caco-2 and Caco-2-SF cells. Their morphological appearance was identical. Slight differences were found in cell growth and expression of brush border enzymes between Caco-2 and Caco-2-SF cells. Similar levels of activity of Gly-Gly transport were expressed in both types of cell. Caco-2 cells cultured on permeable filters showed high transepithelial electrical resistance (TEER), indicating the high monolayer integrity. The transepithelial transport activity for glucose, alanine and Gly-Gly was detected by measuring the change in short-circuit current (ΔIsc) after adding each of these nutrients to the apical chamber. In Caco-2-SF cells, such parameters as TEER and ΔIsc were reduced drastically, suggesting that the monolayer integrity and cell polarity that are important for transepithelial transport were not attained. These parameters, however, could be restored by adding FCS or by milk whey. The result suggested that FCS and milk whey contain factors which regulate the formation of the tight junctions and, consequently, the development of cell polarity. Thus the Caco-2-SF cell-culture system will provide a useful model for studying factors which regulate the intestinal transepithelial transport functions.

Key words

Caco-2 epithelial monolayer human intestine peptide transport serum-free culture 

Abbreviations

BCECF

2′,7′-bis(carboxyethyl)-5(6)-carboxyfluorescein

TEER

transepithelial electrical resistance

LY

lucifer yellow CH lithium salt

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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Kei Hashimoto
    • 1
  • Makoto Shimizu
    • 2
  1. 1.School of Food and Nutritional SciencesUniversity of ShizuokaShizuokaJapan
  2. 2.Department of Agricultural ChemistryThe University of TokyoBunkyo-ku, TokyoJapan

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