The black soldier fly (Hermetia illucens) is attracting increasing interest for its ability to convert low-value substrates into highly nutritious feed. This study aimed at evaluating grape pomace from two varieties (Becuet – B; Moscato – M) as rearing substrates for black soldier fly larvae (BSFL), focusing on the related effects on larval growth performance, proximate composition, and fatty acid profile. A total of six replicates per treatment, and 1 000 BSFL per replica, were used. Larval development was assessed by larvae weight, which was recorded eight times during the trial: the day after the beginning of the trial, and then on days 5, 8, 13, 15, 20, 22, and 27 (day in which the 30% of BSFL reached the prepupal stage). Production and waste reduction efficiency parameters, namely the growth rate, substrate reduction and substrate reduction index, were calculated. The two grape pomace varieties were analysed for their proximate composition and fatty acid profile; the same analyses were conducted on BSFL (30 larvae per replica) that were collected at the end of the trial (day 27). The growth rate of BSFL showed a higher value when the larvae were reared on B substrate (4.4 and 3.2 mg/day for B and M, respectively; P < 0.01). The rearing substrate did not significantly affect the proximate composition of BSFL. The percentage of total lipids (TL) in M-fed BSFL was significantly higher than in B ones. Total saturated (P < 0.001) and monounsaturated fatty acids (P < 0.05) were significantly higher in M-fed BSFL, while an opposite trend was observed for total branched-chain (P < 0.001) and total polyunsaturated fatty acids (P < 0.001). Interestingly, some conjugated linoleic acid (CLA) isomers [i.e., C18:2 c9t11(+t7c9+t8c10) and t9t11] were detected in low amounts in both rearing substrates (total CLA equal to 0.085 and 0.16 g/100 g TL in B and M substrate, respectively). Some CLA isomers (i.e., C18:2 c9t11, t7c9, and t10c12) were also found in BSFL, reaching a total CLA concentration equal to 2.95 and 0.052 g/100 g of TL in B-fed and M-fed BSFL, respectively. This study demonstrates that winery by-products from different grape varieties can significantly affect the development and lipid composition of BSFL. The CLA biosynthesis potential of BSFL opens newsworthy perspectives for a new valorisation of winery by-products to produce full-fat black soldier fly meal and black soldier fly oil enriched in specific fatty acids of potential health-promoting interest.