To investigate the efficacy of umbilical cord serum eyedrops for the treatment of neurotrophic keratitis. Prospective noncomparative case series. Twenty-eight eyes of 28 patients with neurotrophic keratitis who were refractory to conventional treatment. The patients with neurotrophic keraitis were treated with 20% umbilical cord serum eyedrops 6 to 10 times a day. Ophthalmic examinations including best-corrected visual acuity (VA) measurement, corneal sensitivity test, corneal fluorescein staining, and anterior segment photography were performed before and after the treatment. Concentrations of substance P, insulinlike growth factor 1 (IGF-1), and nerve growth factor (NGF) in umbilical cord serum, normal peripheral blood serum, and tears were measured. Epithelial healing time; changes of VA and corneal sensitivity after treatment; and levels of substance P, IGF-1, and NGF in umbilical cord serum, normal peripheral blood serum, and tears. The epithelial defect healed completely in all eyes, with a mean healing time of 4.4+/-4.0 weeks. The epithelial defect healed within 2 weeks in 8 eyes (28.6%), between 2 and 4 weeks in 14 eyes (50.0%), and after 4 weeks in 6 eyes (21.4%). After treatment, VA improved by >2 lines in 17 eyes (60.7%). Mean pretreatment corneal sensitivity was 21.1+/-10.5 mm, and mean posttreatment corneal sensitivity was 24.3+/-11.7 mm (P<0.01). Mean concentrations of substance P, IGF-1, and NGF were 245.3+/-53.9 pg/ml, 239.0+/-77.1 ng/ml, and 729.7+/-72.0 pg/ml in umbilical cord serum; 169.5+/-81.0 pg/ml, 375.5+/-51.3 ng/ml, and 401.7+/-98.1 pg/ml in peripheral blood serum; and 69.8+/-24.9 pg/ml, 75.7+/-50.5 ng/ml, and 107.5+/-70.9 pg/ml in tears, respectively. Umbilical cord serum contains many neurotrophic factors, and umbilical cord serum eyedrops appeared to be effective for the treatment of neurotrophic keratitis.