Towards The Use of Endometrial Mesenchymal Stem Cells as A Cell-based Therapy

We are using a tissue engineering approach to address problemsassociated with using vaginal mesh for treating POP. Wepropose to use autologous endometrial mesenchymal stem cells(eMSC) delivered on polyamide/gelatin composite meshes(eMSC/PA+G) to improve mesh biocompatibility and regeneratevaginal tissue damaged from childbirth injury.eMSC are purified from biopsies obtained from premenopausaland short-term estrogen-treated post-menopausal women usingSUSD۲ magnetic-bead sorting. eMSC are culture-expandedin serum-free medium under hypoxia with a small moleculeTGFβ-receptor inhibitor, A۸۳-۰۱, generating ۹۰-۹۵% SUSD۲+eMSC. ATACseq and RNAseq revealed that A۸۳-۰۱ maintainseMSC stemness by opening chromatin loci enriched for transcriptionfactor binding sites and upregulating gene networksinvolved in developmental and stem cell signalling pathways.A rat model treated with human eMSC/PA+G constructs showedthat eMSC increased vascularisation, reduced chronic inflammation,promoted deposition of crimped collagen, generatinga biomechanically less stiff mesh/tissue complex compared toPA+G. The eMSC had a paracrine mechanism of action in improvingbiocompatibility of PA+G mesh.We are using an autologous preclinical ovine model of vaginalsurgical repair in multiparous ewes. POP is assessed using amodified POP-Q and our novel fibre-optic pressure sensor device.Ovine eMSC are isolated from hysterectomies by FACSsorting CD۲۷۱+CD۴۹f- cells, culture-expanded and labelledwith IODEX-FITC paramagnetic nanoparticles for cell trackingbefore seeding on PA+G mesh and implanting transvaginallyusing urogynaecological surgical procedures.Our tissue engineering approach using autologous eMSC deliveredon polyamide/gelatin scaffolds addresses some problemsassociated with the use of vaginal mesh and may improve surgicaloutcomes for treating POP.