Background: Globally, reproductive aging is increasing and is inextricably linked to aging in general. Premature ovarian insufficiency (POI), which has primarily had an idiopathic etiology up to this point, is an extreme manifestation of reproductive aging. Aim: was to evaluate the effectiveness of instructional guidelines on awareness and perception of premature ovarian insufficiency women regarding stem cell-based ovarian regeneration. Methods: A quasi-experimental one group pre-test/post-test design was conducted at Menoufia University Hospital with 152 women diagnosed with POI. Women completed a structured interviewing questionnaire, an awareness assessment questionnaire, and a perception assessment scale. Results: Post-intervention, awareness levels and positive perception of stem cell therapy significantly improved (p < 0.001). Conclusion: Instructional guidelines effectively enhanced awareness and perception of POI women regarding stem cell-based ovarian regeneration. Recommendation: These findings highlight the need for continued education and improved healthcare accessibility to support informed decision-making on emerging fertility treatments.