Our study reports the triglyceride (triacylglycerol, TAG) composition of five new Tunisian virgin olive oil cultivars obtained through controlled crossings of the cultivar (cv.) Chemlali Sfax. These cultivars have been selected among a progeny of 500 olive descendants, based on an evaluation of the fatty-acid (FA) composition of their oils. Among these samples, two were derived from the crossing with the cv. Sigoise as pollinator (SM634) or pollen acceptor (SM1110) and the others from the crossing with the cv. Meski as pollen acceptor (SM513, SM514, and SM517). The five descendants were characterized by a good fat value, a balanced FA composition, and a high content of triolein, varying between 26.9 (SM514) and 45.46% (SM1110). They had an improved FA composition as compared to that of the cv. Chemlali Sfax and their fruits were slightly bigger. The principal component analysis suggested that the TAG variables were more suitable than the total FAs for an optimum classification of the cultivar samples analyzed. The cultivars obtained through the crossing with the cv. Sigoise (as pollen acceptor or pollinator) had a more favorable composition of FAs and TAG than those obtained through the crossing with the cv. Meski, which indicated that genetic factors had the most important influence on the quality of the virgin olive oils.