Differentiation of human endometrial stem cells into germ cell – Like cell in fibrin scaffold

Abstract

Recent studies on stem cells differentiation into germ cells have changed scientists’ attitude to reproductive problems as well as infertility topics. It is supposed there are promising and new approaches in treatment of infertile couples and numerous advances will be made in reproductive medicine in near future. Application of embryonic stem cells for clinical trials is limited due to high potent of tumorogenicity and ethical issues. Therefore, pluripotent cells taken from adult tissues or organs, could be a good alternative for gamete production. Herein, we hypothesize to stimulate human endometrial stem cells (hEnSCs) differentiation into germ cell-like cells by culturing in retinoic acid (RA) as 2D medium and then in fibrin as 3D scaffold. Germ cell markers such as DAZL, DDX4 and Dppa3, will be assessed by immunofluorescence and real-time PCR. Fibrin mechanical properties will be examined by rheology analysis and cell viability will be determined by MTT assay. Specific markers expression and the cells’ integrity will be detected by immunofluorescence staining and SEM analysis respectively. We suggest differentiation of hEnSCs into germ cell-like cells in a medium containing 10⁻⁵ M RA in which the specific markers were expressed properly in both 2D and 3D medium cultures. Additionally, fibrin scaffold will offer a proper 3D scaffold for hEnSCs-derived germ cell-like cells.