Purpose: To assess patient satisfaction with autologous serum tears (AST) for the treatment of dry eye disease (DED). Methods: A prospective, observational case-series, including 100 participants who were treated with AST for DED. The five-item Dry Eye Questionnaire (DEQ-5), Symptom Assessment in Dry Eye (SANDE) Questionnaire, and the Treatment Satisfaction Questionnaire for Medication (TSQM) were used to assess DED symptoms and patients' treatment satisfaction. Results: The duration of AST treatment was 1-6 months in 25%, 6-12 months in 24%, 1-2 years in 22%, and 2-3 years in 29% of the participants. The mean cumulative DEQ-5 score was 12.7 +/- 3.8 and 65% of the participants scored >= 12, which may suggest an underlying diagnosis of Sjogren's syndrome. The use of other treatments was significantly reduced after the use of AST (P < 0.001). The mean total SANDE score improved from 79.5 +/- 21.8, 95% CI [75.2, 83.8] before to 42.2 +/- 23.2, 95% CI [37.6, 46.8] after using AST (P < 0.001). The mean TSQM treatment efficacy, convenience, and global satisfaction scores were 73.1 +/- 17.2, 95% CI [69.6, 76.5], 72.5 +/- 17.5, 95% CI [69.0, 76.0], and 74.6 +/- 22.7, 95% CI [70.1, 79.0]. No patients reported any side effects. Conclusion: Autologous serum tears treatment in severe dry eye disease significantly reduced dry eye symptoms, with high patients' satisfaction scores.