Cycling infrastructure has emerged as a strategic lever for sustainable urban development in Latin America, yet the factors that determine its effectiveness remain poorly understood and dispersed across fragmented literature. This systematic review, conducted in accordance with PRISMA 2020 guidelines, synthesizes evidence from 16 peer-reviewed studies published between 2020 and January 2026 to identify the key determinants of cycling infrastructure effectiveness in Latin American cities. Five critical dimensions emerged from the analysis: physically segregated infrastructure, gender-responsive design, multimodal connectivity, environmental and public health co-benefits, and adaptation to topographical and climatic conditions. The evidence reveals that segregated bike lanes increase cycling uptake by 48–187%, multimodal integration boosts combined trips by approximately 34%, and gender-sensitive interventions increase female cycling participation by up to 89%. These findings converge on a central insight: infrastructure effectiveness is not achieved through isolated investments but through the coherent, simultaneous implementation of multiple interconnected dimensions. This review contributes an original multidimensional analytical framework to guide evidence-based public policy and investment in sustainable urban mobility across the region.