Objectives: As part of the regionalization of public health initiatives, our objective was to generate and compare regional data on antibiotic resistance in Escherichia coli isolated from urban settings. Method: We analyzed the regional frequency of acquired antibiotic resistance in E. coli routinely isolated in 2013 from the urine of outpatients using data from a national network of private medical laboratories. Results: A total of 51,463 E. coli strains from 11 of the 13 new French metropolitan regions were analyzed. The overall proportion of extended-spectrum beta-lactamase (ESBL)-producing strains was 3.3% and ranged from 1.8% to 5.1% across regions. Among the 6 antibiotics evaluated, the overall resistance rate was lowest for furans and fosfomycin (1.3% for both) and highest for co-amoxiclav (33.9%) and cotrimoxazole (20.4%). The difference between the region with the highest resistance rate and the one with the lowest was 1.1% for furans and 2.9% for fosfomycin. Overall resistance to at least 3 of the 6 antibiotics tested (multidrug resistance) was 6.2%. It ranged from 5.0% to 10.2% depending on the region. Conclusion: The establishment of a national surveillance network for resistance in urban areas has made it possible to assess regional differences in the prevalence of antibiotic resistance in E. coli. Further studies will need to be conducted to try to explain some of these differences. Strengthening the network would be necessary to ensure its long-term viability and to fill in the missing data.