Improving reproductive efficiency is essential for meeting the growing demands of livestock production. In vitro production (IVP) of bovine embryos has enormous potential for enhancing reproductive efficiency of elite animals. However, its efficiency remains low due to poor rates of embryo development and reduced cryosurvival of IVP embryos, necessitating further research. The objective was to compare effects of various maturation media components, including fetal bovine serum (FBS), FBS + L-carnitine, and bovine serum albumin (BSA) on oocyte maturation, lipid content, developmental competence, and cryotolerance of resulting embryos. Additionally, impacts of L-carnitine in FBS-containing media on Hippo signaling in bovine oocytes was also examined. Bovine ovaries were collected from a local abattoir and oocytes were recovered and matured in vitro with FBS, BSA, or FBS + 1.5- or 3.0-mM L-carnitine. Oocyte maturation was assessed based on polar body formation, lipid content was assessed with Nile Red staining, and embryo development was evaluated by assessing cleavage and blastocyst rates. Embryos were cryopreserved and cryotolerance was assessed by evaluating their ability to undergo re-expansion and hatching. Expression of Hippo pathway genes and maturation markers were assessed through RT-qPCR. Maturation rates did not differ among FBS alone (83.2%) and FBS with L-carnitine (1.5 or 3.0 mM; 82.0 and 80.0%, respectively). However, the BSA group had a lower maturation rate (70.6%) compared to other groups. Expression of maturation markers, cyclin B and Cdk1, were similar between FBS and FBS + L-carnitine groups. Lipid content was reduced significantly in BSA and FBS + L-carnitine compared to the FBS. Neither cleavage nor blastocyst rates differed between FBS and FBS + L-carnitine groups. Oocytes matured with 1.5 mM L-carnitine had a significantly higher cleavage rate compared to BSA. Post-thaw, embryos from the FBS + L-carnitine group had the highest re-expansion rate at 24 h (78.8%), whereas the BSA and FBS groups had lower rates. Expression of Hippo pathway genes was not significantly altered by L-carnitine. Therefore, it was concluded that L-carnitine enhanced cryotolerance and reduced lipid accumulation without affecting maturation or Hippo signaling.