Background Current literature highlights a gap in precise stroke cost data for Latin America. This study measures the real costs associated with acute ischemic stroke care in Latin America using Time-Driven Activity-Based Costing (TDABC). The findings aim to lay a solid foundation for adopting value-based healthcare (VBHC) strategies in the region. Methods The study is an observational, multicenter, international analysis of direct costs and outcomes for patients hospitalised with acute ischemic stroke from December 2021 to December 2022. Data from stroke centres in Argentina, Brazil, Chile, Colombia, Costa Rica, Mexico, Peru, and Uruguay were analysed. Costs were stratified by country. Factors such as favourable outcomes based on the modified Rankin Scale (mRS 0-2), clinical risk levels, and treatment interventions were considered for the analysis. Generalized Estimating Equation (GEE) models were utilised to assess the relationship of clinical variables with the total cost per patient. Findings A total of 1106 patients were included in the study. Among these patients, 74% received medical treatment alone, 18% received intravenous thrombolysis (IVT), 4% underwent mechanical thrombectomy (MT), and 3% received combined IVT plus MT. The mean cost per patient was I$ 12,203 (SD I$ 15,055), with 49% achieving a favourable functional outcome. Compared to medical treatment alone, MT incurred costs 3.1 times higher, with an incremental cost of I$ 20,418 per patient (p < 0.0001). Across all countries, costs increased according to patients' clinical risk and treatment options, with length of hospital stay emerging as the primary cost driver. Interpretation Our study highlights significant disparities in stroke costs across healthcare services in Latin America, influenced by variations in treatment accessibility, patient outcomes, and clinical risk profiles. These findings offer essential insights for shaping health policy decisions to enhance the long-term sustainability of stroke care in the region. Copyright (c) 2024 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).