To date, there is no compelling evidence of the optimal timing of surgery for congenital diaphragmatic hernia (CDH). This study aimed to establish the optimal timing of surgery in neonates with isolated left-sided CDH. This multicenter cohort study enrolled 276 patients with isolated left-sided CDH at 15 institutions. Based on the timing of surgery, the patients were classified into four groups [< 24h (G1), 24-47h (G2), 48-71h (G3), and ≥ 72h (G4)]. The clinical outcomes were compared among the groups using a logistic regression model, after adjusting for potential confounders, such as disease severity. Multivariate analyses showed that G2 also had a lower mortality rate than the other groups. In mild and severe cases, there were no significant differences in mortality across the four groups. In moderate cases, G2 resulted in significantly increased survival rates, compared with G1. The study results suggest that surgery within 24h of birth is not recommended for patients with moderate severity CDH, that there is no benefit in the delay of surgery for more than 72h in mild severity CDH patients, and that there is no definite optimal time to perform surgery in severe cases of CDH.