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Since ancient times, thistles have been used as clotting agents in the production of traditional cheeses, particularly in the Mediterranean area. In recent years, their use in cheesemaking has increased to satisfy the growing requests from vegetarian consumers. In this research paper, four different cheeses, typical of the Mediterranean area, were evaluated from a nutritional point of view: Caciofiore (from Italy) and Torta del Casar (from Spain), both typically produced using vegetable rennet, and Queso de Murcia al vino (from Spain) and Feta (from Greece), traditionally produced using animal rennet. All the cheeses were manufactured according to their traditional cheesemaking procedures and used as controls. Experimental cheeses were produced using aqueous extracts obtained from flowers of either spontaneous or cultivated thistles indigenous to the Mediterranean area (respectively Onopordum tauricum for Caciofiore, and Cynara humilis for Torta del Casar, Queso de Murcia al vino, and Feta). All cheeses were characterized for fat-soluble and mineral compounds to assess their nutritional adequacy according to the recommended daily intake of each evaluated nutrient. All the cheeses were found to be a good source of vitamin A, calcium and phosphorus, with an optimal Ca/P molar ratio, except for Feta. By consuming the recommended serving (50 g) of the studied cheeses, the salt and cholesterol intake is, on average, 16.4% and 15.9%, respectively of recommended intake. The use of aqueous thistle extracts in cheesemaking appears to have no effect on the nutritional quality of the studied cheeses.