Background: The CO{}_2 laser has been used to treat vulvar lichen sclerosus (VLS) with a significant reduction in symptoms, and there is a paucity data on the efficacy, sexual function, and quality of life (QOL) improved. This study was to evaluate the efficacy of fractional CO{}_2 laser for VLS and assess sexual function and QOL from multi-centers. Methods: The women with biopsy-proved VLS who were treated with CO{}_2 fractional laser were enrolled between January 2017 and December 2018 from three centers. The visual analogue scale (VAS) was to assess clinical symptoms. The Female sexual distress scale-revised (FSDS) was to evaluate the patients’ sexual activity, and the family Dermatology Life Quality Index (DLQI) was to evaluate the QOL. Results: A total of 119/122 patients (mean 44.5 years, 27–72 years) completed the treatment with fractional CO{}_2 laser and following-up. The clinical symptoms and QOL were improved significantly from baseline to follow-up (3 months and 12 months post-treatment) according to the following measures: itching score, 7.65 (1.07, 0–10) versus 4.52 (1.23, 0–10) versus 0.96 (1.49, 0–4); burning score, 6.9 (1.66, 0–10) versus 3.82 (1.79, 0–8) versus 0.98 (1.42, 0–4); DLQL score, 14.24 (5.64, 0–30) versus 8.7 (1.9, 0–15) versus 5.7 (2.3, 0–10); FSDS score, 15.48 (11.3, 0–51) versus 12.51 (10.59, 0–45) versus 7.87 (9.34, 0–50). Conclusion: In conclusion, the fractional CO{}_2laser was effective and safe for treating VLS, which significantly ameliorated the annoying symptoms and improved the quality of life.