Journal
ALTEX-ALTERNATIVES TO ANIMAL EXPERIMENTATION
Volume 30, Issue 2, Pages 159-168Publisher
SPEKTRUM AKADEMISCHER VERLAG-SPRINGER-VERLAG GMBH
DOI: 10.14573/altex.2013.2.159
Keywords
alternative methods; intestinal permeability; brush border enzymes; MDR-1/P-glycoprotein; peptide transporter-1; serum-free medium
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Funding
- EU FP6 Project LIINTOP [STREP-037499]
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Human intestinal Caco-2 cells were differentiated using serum-reduced medium with fetal bovine serum (FBS) added only to the basolateral (BL) medium, and four serum-free media, containing insulin, transferrin, selenium (ITS), or MITO+(TM) serum extender (ITS plus growth factors), with or without addition of a lipid mixture, respectively. Differentiation was assessed by monitoring monolayer permeability, alkaline phosphatase and sucrase activities, and the transport of digoxin and cephalexin. Notably, the serum-reduced protocol produced results that were comparable to cells differentiated in the control medium and should be recommended as an alternative to the use of 10% FBS in both apical (AP) and BL media. ITS serum-free medium elicited permeability values and cephalexin transport similar to control cells. MITO+(TM) medium was the most efficient in promoting the two transport activities investigated, and it should be further evaluated with a larger set of substances, although its undisclosed composition represents a limit that may override these advantages.
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