Purpose The aim of this study was to evaluate the role of leg alignment on long‐term clinical outcome after matrix‐associated autologous chondrocyte implantation (M‐ACI) and to define an individualized target range to optimize clinical outcome. Methods The present study examined patients who underwent M‐ACI of the femoral condyle. The Knee Injury and Osteoarthritis Outcome Score (KOOS) and Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) were used to assess the results. Clinical outcomes were related to Patient Acceptable Symptomatic State (PASS). For intra‐ and interobserver reliability of mechanical tibiofemoral angle, mechanical medial proximal tibial angle and mechanical lateral distal femoral angle, we calculated intraclass correlation coefficients using a two‐way mixed model with absolute agreement. A regression model and receiver‐operating characteristics curve were used to identify an individual range of alignment where a favourable clinical outcome could be expected in the long term. Results Additional osteotomy was performed in 50% of patients with similar clinical outcomes as physiologically aligned patients (p > 0.05). The curve‐fitting regression model identified a target range of −2.5° valgus to 4.5° varus for ideal postoperative alignment (R2 = 0.12, p = 0.01). Patients within this range were more likely to achieve PASS (70% vs. 27%, p = 0.001). In medially treated defects, a refined range of −2.5° valgus to 4° varus alignment was found (R2 = 0.15, p = 0.01). These patients were more likely to achieve PASS (67% vs. 30%, p = 0.01) and showed favourable postoperative KOOS and MOCART scores (p = 0.02). Patients with lateral defects were more likely to achieve PASS within a range of −2° valgus and 0.5° varus (90% vs. 45%, p = 0.03) and showed favourable postoperative KOOS and MOCART scores (p = not significant). Conclusions An individual range of leg alignment—whether achieved by osteotomy or physiologic alignment—should be respected in M‐ACI treatment. A neutral to slightly undercorrected alignment favours the postoperative outcome after M‐ACI. When planning surgery for patients with focal cartilage defects of the femoral condyle, these ranges should be recognized as critical factors. Level of Evidence Level III.