The study was conducted to determine the content and retention of selected fat-soluble vitamins and minerals in curd cheese–tvarog made by a traditional method, taking into account the effect of the quality of the raw milk and the type of starter cultures used. The raw milk used to make the tvarog was obtained in various conditions, i.e., with and without the use of pasture forage (in a traditional and an intensive system), in two production seasons (spring/summer and autumn/winter), from two breeds raised in Poland (the Black-and-White variety of Polish Holstein-Friesian and Simmental). Two variants of starter cultures were used to make tvarog: Freeze-dried DVS starters (Flora Danica) and pure cultures of mesophilic lactic acid bacteria. The acidity and content of protein, fat, selected fat-soluble vitamins (A, D3, and E), and selected macro-elements (Ca and Mg) were determined in samples of bulk milk and cheese. Retention rates of individual nutrients from the milk to the cheese were calculated. A higher content of fat-soluble vitamins was found in milk obtained from Simmental cows kept in a traditional system in the spring/summer season, as well as in the tvarog produced from it. Vitamin retention rates from the raw material to the tvarog were above 90%. The mineral composition of the cheese was not associated with the quality of the milk used. Very low retention rates from milk to cheese were obtained for Ca and Mg (below 20%). Higher retention rates were obtained in the spring/summer season when culture variant 1 was used. However, the starter culture was not found to significantly influence the concentration or retention of vitamins in the experimental cheese.