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Foods and supplements with added phytosterols are widely recommended as dietary means to lower LDL-cholesterol, one of the key cardiovascular risk factors. Phytosterols interfere with cholesterol absorption from both the diet and endogenous sources. The behaviour of phytosterol-formulations in a gastrointestinal environment is important to predict their efficacy in vivo. In vitro models can serve as screening tool to guide product development and to facilitate meeting consumer demand for convenient new formats that best fit modern lifestyles.