The search for functional foods or functional food ingredients, i.e. foods or food ingredients that can enhance health, is beyond any doubt one of the leading trends in today's food industry. In this context, probiotics, i.e. living microbial food supplements, and prebiotics, i.e. non-digestible food ingredients, receive much attention. Both popular concepts target the gastrointestinal microbiota. While in the Western world, intake of probiotics has been recommended for long, prebiotics in general, and non-digestible oligosaccharides in particular, have only recently received attention. This review deals with production and characterization of non-digestible oligosaccharides and focuses on their role in promoting health and treating diseases. Attention is paid to the effects of non-digestible oligosaccharides on constipation, mineral absorption, lipid metabolism, cancer prevention, hepatic encephalopathy, glycemia/insulinemia, and immunomodulation.