M Haghshenas - Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran. Department of Developmental Biology, Faculty of Basic Scienc-es and Advanced Technologies in Biology, University of Scie
S TAVANA - Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
LEILA MONTAZERI - Department of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
E ZAND - Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

Background: 3D culture of ovarian follicles using natural hy-drogel-based scaffolds to attain fertilizable oocytes is consid-ered as a promising strategy in women suffering from infertility. Human amniotic membrane due to its unique biological com-position and regenerative properties, has been used extensively in tissue engineering. In this study, mouse secondary follicles were cultured using decellularized human amniotic membrane (DHAM) hydrogel in order to improve ovarian follicle culture. Materials and Methods: Ovarian follicles (120-140µm) were cultured in three different doses of DHAM hydrogel (15, 30 and 45 mg/ml) in combination with alginate (1.5%) (1:1) and com-pared to control group (Alginate 0.75%). At the last days of the culture period (14 days), follicular morphology was evaluated. Results: The results have been showed that antral/total rate of follicles in 45 mg/ml of DHAM hydrogel (42.77%) were sig-nificantly (P<0.05) higher than the other dozes but there wasn’t any significant difference between 45mg/ml and control one (46.08%). Also, the rate of degenerated follicles in 45mg/ml (51.85%) reached to the control group (42.62%) and was the lowest rate as compared to the other hydrogel-alginate com-binations in which was comparable against 30 mg/ml group (68.69%).Conclusion: DHAM-based hydrogel as a bioscaffold may ap-ply instead of alginate which can improve the rate of antral follicles and decrease the rate of degenerated follicles in 3D culture system.

Secondary Follicles, 3D Culture, Human Amniotic Membrane, Mouse