Abstract
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+™ 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+™ 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.
Highlights
No major increase in alkaline phosphatase (ALP) activity was observed between days 15 and 21 in Caco-2 cells differentiated in all experimental media (Fig. 2A)
On day 15 and day 21 cells differentiated in fetal bovine serum (FBS) Asymmetric medium showed significantly higher values, while cells differentiated in all other serum-free media showed significantly lower values compared to cells in FBS Symmetric control medium
Cells differentiated in FBS Asymmetric medium showed higher values of SUC activity at both days 15 and 21, which was significantly different from activity of cells in FBS Symmetric medium
Summary
The aim of the present study was to investigate the effects of distinct serum-free or serum-reduced media on the differentiation of Caco-2 cells. The purpose of this work was to provide recommendations for the use of serum-free or serum-reduced protocols for differentiation of Caco-2 cell monolayers, with the ultimate aim of contributing to the reduction in the use of serum as culture medium supplement
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