Tissue Engineered Human Amniotic Membrane Application in Mouse Ovarian Follicular Culture

Since folliculogenesis requires a powerful cell-matrix interaction, natural scaffolds seem to be needed for follicular culture. Human amniotic membrane (HAM) offers promise as a support of in vitro ovarian follicular culture. HAM was decellularized with trypsin and EDTA. DNA and histology assays were performed to determine the elimination rate of genomic components. Cyto-biocompatibility of decellular AM (DAM) was verified by the cell viability (MTT) test. The small parts of intact amniotic membrane (IAM) and DAM were coated on the bottom of 96-well and each well was filled with 150 µL of base medium. Mouse primary-secondary (PS) follicles were separated to three groups: 1-culture in base medium (Control), 2-culture on IAM and 3-culture on DAM. Follicular size, morphology, viability, estradiol production and genes expression were evaluated and IAM group showed better growth and development in follicle culture. The viability rate and estradiol production in both experimental groups were statistically higher than the Control. Gdf9, Bmp15 and Cx37 were found to have higher expression levels in IAM group. Also, maximum apoptotic and survival indexes were determined in Control and IAM groups, respectively. Finally, IAM provides a better protective environment for mouse PS follicular culture that can reduce apoptosis level.