In Vitro Model Three-Dimensional Culture for The Study of Human Implantation

Background: Implantation involves coordinated interactionbetween the embryo and the receptive endometrium. Implantationfailure is one of the major causes of subfertility. However,the detail mechanism of implantation is still not very clear.Studying human embryo implantation is challenging since invivo experiments are impractical and unethical, and studies inanimal models do not always translate well to humans. Thus,it is necessary to develop an in vitro experimental model thatcan better reflect the in vivo situation for studying implantation.Materials and Methods: In vitro novel three-dimensional (۳-D) endometrium cultures were constructed with fibrin-agaroseas matrix scaffold. Endometrial biopsies were obtained fromhealthy, fertile women on cycle day LH+۴ and stromal andepithelial cells were isolated. Stromal and epithelial cells werecharacterized by flow cytometry and immunocytochemistry.Human endometrial stromal cells embedded in a mixture offibrin-collagen I that covered with a layer of epithelial cells andcultured until confluence. Healthy, viable human embryos wereplaced on cultures. Two days later, the cultures were tested forthe development and attachment of embryos. The biologicalgrowth, live and dead staining, MTT assay and haematoxylinand eosin staining of cultured cells were performed.Results: Histological staining showed that epithelial cells wereorganized in a polarized columnar epithelium on the apical surfacegel. A polymer gel that not only supports cell growth buthandily transforms from gel to liquid After a few days. Cellviability was evaluated using live/dead and MTT assay showedthat the ۳D cultures were not toxic for the cell. Cell viabilitytests showed that stromal and epithelial cells developed andsurvived and were compatible with scaffold.Conclusion: In this study, an in vitro-three dimensional embryo-endometrial cell culture model to investigate the humanembryo implantation process was established that appeared toimitate of the normal human endometrium.

کلیدواژه ها:

Three-Dimensional Culture ، Implantation ، Human Endometrium ، Model