Establishment and Characterization of Vaginal Tissue Primary Culture: Feasibility of Cell Therapy for Mayer-Rokitansky-Kuster-Hauser Syndrome (MRKHS) Patient Treatment

Abstract

Title: Establishment and Characterization of Vaginal Tissue Primary Culture: Feasibility of Cell Therapy Usage in Mayer- Rokitansky-Kuster-Hauser Syndrome (MRKHS) Patient´s Treatment. Background: The uterine and vaginal agenesis, Mayer- Rokitansky-Küster-Hauser Syndrome (SMRKH), is a congenital malformation that implies in the impairment of sexual activity and reproductive life. The first line therapy is nonsurgical (dilatation method) but surgical methods may be necessary to create a functional neovagina. In the surgical procedure, various types of graft materials have been used to line the neovagina, and recently, alternative grafts have been studied to minimize adverse effects. Our aim was to establish vaginal primary cultures in order to prove the feasibility of cell therapy for the reconstruction of the vaginal lining. Methods and findings: Nine small biopsies were processed according to explant technique in order to isolate vaginal cells (VC). Different cell types were obtained, including fibroblast and epithelial cells, according to optical microscopy, flow cytometry and immunofluorescence assessment. These cells could not differentiate into mesenchymal lineages (adipogenic, chondrogenic and osteogenic) but are able to form tissue-like cell aggregates. The expression of telomeres and telomerase genes were similar to the endometrial fibroblast. Conclusion: We are able to obtain and characterize vaginaderived cells in primary cultures. The isolated cells seem to be good candidates to cell therapy usage in MRKHS as they have phenotypic and physiological characteristics suitable to the reconstruction of vaginal lining.