Human endometrial stem cells (hEnSCs) differentiation into germ cell-like cells by encapsulating in fibrin scaffold
Maryam Roya Ramzgouyan, Seyed Mohammad Tvangar, Jamshid Hajati, Nasrin Lotfibakhshaiesh, Fardin Amidi,
Somayeh Ebrahimi-Barough, Roya Karimi, Reza Roozafzoon, Mina Jafarabadi, Ahmad Jafar Bekloo, Jafar Ai
Journal Title:Basic Research Journal of Medicine and Clinical Science
In recent years, stem cells differentiation into germ cells has focused on infertility topics. A good alternative for gamete production in clinical trials would be pluripotent cells isolated from adult tissues. Herein, we identified the suitable conditions which promoted human endometrial stem cells (hEnSCs) differentiation into germ cell-like cells by adjusting the best concentration of retinoic acid (RA) in 2D medium and cultured them in fibrin as a 3D scaffold afterwards. The optimum concentration of RA was 10-5 M for differentiation after 7 days. Characteristic cell markers such as DAZL, DDX4 and Dppa3 were assessed by immunofluorescence and real-time PCR. The mechanical properties of the fibrin were examined by rheology analysis. After encapsulating the hEnSCs-derived germ cell-like cells in fibrin hydrogel, cell viability, specific markers expression and the cells' integrity in fibrin were assessed by using MTT assay, immunofluorescence staining and SEM analysis respectively. We propose that hEnSCs can be differentiated 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. In addition, fibrin hydrogel could offer a proper 3D scaffold for hEnSCs-derived germ cell-like cells regarding tissue engineering of gonads.
Keywords: Human Endometrial Stem Cells (hEnSCs), Differentiation, Retinoic Acid (RA), Fibrin Hydrogel, Germ Cells