Objective: To investigate the comprehensiveness, quality, and reliability of YouTube videos that target teaching patients gonadotropin self-injections. Study design: Videos demonstrating gonadotropin self-injections were searched using the keywords “Gonal-f injection,” “Puregon injection,” “Menapur injection,” “Merional injection,” “Fostimon injection,” and “Menagon injection,” on December 20th, 2020. The videos were divided into two categories as including “useful information” and “misleading information” by two physicians. A 5-point global quality scale (GQS) and 5-point modified DISCERN scale were used for the assessments of quality and reliability, respectively. Results: Among 110 videos, 90 (81.8%) were found to include useful information and 20 (18.2%) were found to give misleading information. The kappa statistic for inter-observer agreement was 0.817 (p < 0.001). Useful videos were the most comprehensive and had the highest reliability and quality scores. We found that all videos uploaded by universities or professional organizations included useful information. However, there was no significant difference between useful and misleading videos regarding audience interaction analysis parameters (p > 0.05). On the other hand, mean reliability, GQS, and comprehensiveness scores were higher in the useful information group than in the other group. As the subgroup analysis was performed by source, patient opinion videos had lower reliability, comprehensiveness, and GQS scores than videos created by other sources (p < 0.001). Conclusion: Our analysis revealed that there were a significant number of English-language YouTube videos, with high quality, rich content, and reliability that could be sources of information on the accurate technique of gonadotropin self-injections. However, some misleading information videos may lead to negative outcomes. Therefore, physicians should ensure that online sources are comprehensive and reliable for the use of their patients with infertility. Also, YouTube health videos should be checked for both reliability and ethical standards.