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Posterior fourchette fissures can recur and become chronic despite multiple medical and surgical treatments. We report the case of a recurrent posterior fourchette fissure that resolved after subdermal injection with adipose-derived regenerative cells.A 44-year-old primiparous patient experienced severe dyspareunia for 10 years as a result of a recurrent posterior fourchette fissure that began after a vaginal delivery with episiotomy. She was treated unsuccessfully with topical antibiotics, estrogen, and steroids and ultimately with a perineoplasty at the age of 42 years. Adipose-derived regenerative cells were extracted enzymatically from her adipose tissue and injected subdermally and submucosally beneath the fissure and surrounding area. The fissure completely resolved within 3 months after treatment and has not recurred after more than 1 year of follow-up.Subdermal injection of autologous adipose-derived regenerative cells may be a treatment option for treatment of recurrent fourchette fissures that do not respond to standard medical and surgical therapies.