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Simple Summary: Self-sampling for HPV primary screening is recognized as a viable alternative to clinician-sampling. Its use in an Italian cervical cancer prevention program, focusing on acceptance, ease of use, and follow-up adherence, was evaluated. Self-sampling was offered to 19,327 women aged 30–64, overdue for screening due to the COVID-19 pandemic; they had never or irregularly attended. Results showed 11.5% of women chose self-sampling, more than doubling the participation rate compared to clinician-sampling alone (<5%), showing high acceptance. The return rate of self-samples was high (79.5%), with 1.1% being inadequate, indicating ease of use. HPV positivity was higher in the self-sampled group (13.1%) compared to the 2019 ordinary screening population (9.9%). Compliance with follow-up procedures exceeded 90%, and cervical cancer detection rates were higher (0.9‰) than in the routine screening population (0.4‰). Self-sampling was therefore effective in reaching non-attenders, but clinician-sampling remains important, as 6% of women opted for a clinician appointment. Background/Objectives: Self-sampling is recognized as a viable alternative to clinician-sampling for HPV primary screening. This study aimed to assess, within an Italian organized cervical cancer screening program, the acceptance and ease of use of self-sampling and the adherence to follow-up. The prevalences of HPV infection, cervical dysplasia, and cancer were contextually evaluated. Methods: Electronic records of 19,327 women, 30–64 years-old, residing within the Bologna Local Health Authority territory, who were offered self-sampling as an alternative to clinician-sampling, were retrospectively reviewed. They had never or irregularly attended and were overdue for a screening invitation due to the COVID-19 pandemic. An opt-in approach was adopted, involving local pharmacies for kit delivery and sample collection. Initially, HPV-positive results led to direct referral to colposcopy; later, cytological triage on clinician-samples was provided. Results: Self-sampling reached over twice as many women (11.5%) compared to historical clinician-sampling alone (<5%), showing high acceptance. Additionally, a high screening completion level was observed, with 79.5% of self-samples returned to pharmacies. A low percentage of self-samples resulted in inadequate results (1.1%), suggesting the method's ease of use. HPV-positivity was 13.1%, higher than the 9.9% recorded in the ordinary screening population in 2019 (p < 0.001), the last year before the pandemic. Compliance to both immediate colposcopy and cytology triage exceeded 90% (p = 0.675). The rate of cervical adenocarcinoma was twice as high as in the routinely screened population in 2019 (0.9‰ versus 0.4‰). Finally, 6% of women opted for clinician appointments. Conclusions: Self-sampling proved to be an easy-to-use and effective tool for reaching non-attenders, who are at high risk of cancer. Cytology triage on clinician-samples did not negatively impact follow-up adherence. It seems appropriate to maintain a clinician-collection option even among non-attenders. [ABSTRACT FROM AUTHOR]