Epidemiological and interventional studies have clearly demonstrated the beneficial impact of consuming oat and oat-based products on serum cholesterol and other markers of cardiovascular disease. The cholesterol-lowering effect of oat is thought to be associated with the β-glucan it contains. However, not all food products containing β-glucan seem to lead to the same health outcome. Overall, highly processed β-glucan sources (where the oat tissue is highly disrupted) appear to be less effective at reducing serum cholesterol, but the reasons are not well understood. Therefore, the mechanisms involved still need further clarification. The purpose of this paper is to review current evidence of the cholesterol-lowering effect of oat in the context of the structure and complexity of the oat matrix. The possibility of a synergistic action and interaction between the oat constituents promoting hypocholesterolaemia is also discussed. A review of the literature suggested that for a similar dose of β-glucan, (1) liquid oat-based foods seem to give more consistent, but moderate reductions in cholesterol than semi-solid or solid foods where the results are more variable; (2) the quantity of β-glucan and the molecular weight at expected consumption levels (∼3 g day−1) play a role in cholesterol reduction; and (3) unrefined β-glucan-rich oat-based foods (where some of the plant tissue remains intact) often appear more efficient at lowering cholesterol than purified β-glucan added as an ingredient.