Co-culture Of Mouse Embryos With Human Decidua Monolayer Cells

ObjectiveTo achieve a kind of co-culture system for mouse embryos with human decidua monolayer cells and to evaluate the effects of human decidua generation monolayer cells versus cryo-thawed monolayer cells on mouse embryo development. The purpose of this study is to increase the pregnancy rate in patients with implantation failure by use of a co-culture system for human embryos development on autologous endometria monolayer cells.MethodsFirstly, early pregnant patients underwent a decidua biopsy at induced abortion. Then the tissue was digested enzymatically. These cells were cultured to confluence, released, cryopreserved, thawed, achieved generation and cryo-thawed decidua monolayer cells. Secondly, immunohistochemistry was employed to express ER, PR in the cryo-thawed decidua cells. Thirdly, co-cultured for Kunming mouse 1-cell embryo acquied by controlled ovarian hyperstimulation with generated and cryo-thawed human decidua monolayer cells (marked as group A and B, respectively), and cultured with conventional media as control (group C).Results1 Human decidua cells have the viability in in vitro culture, and easily acquire the ability to be cultured and generated.2 The cryo-thawed decidua monolayer cells can be successfully acquired, and ER, PRwas positively expressed by immunohistochemistry. These cells revealed the characteristics of eutopic endometrium.3 Mouse 1-cell embryos were gained after controlled ovarian hyperstimulation, then co-culture with generated and cryo-thawed decidua cells until blastocyst hatched.4 Co-culture with generated decidua cells can improve the formation of 4-cell stage, there was no the same result in group B and C. But the formation rate of 8-cell stage significantly differ among group A, B and C by x2 procedures, whereas there did not exist between group A and B.5 The morular rate, early blastocyst rate, and expanding blastocyst rate of mouse embryos in group A and B were significantly higher than those of group C. There was no difference between group A and B.6 The formation rate of high quality embryos of groups A and B were significantly higher than those of group C. There was also no difference between group A and B.7 Pathogen was examined in culture medium, bacteria and fungus were not found.Conclusions1 The co-culture system for mouse embryo development on human decidua monolayer cells is a feasible method for analysing the effects and discussing the apoptosis mechanism of co-culture.2 The generated or cryo-thawed human decidua monolayer cells as a kind of feeder cells can improve the mouse embryo development. It can be used as a feasible method for studying human embryo development on autologous post proliferative to early secretory endometrium monolayer cells.4 Contamination can be avoided by strictly aseptic operation. The method of co-culture is safe and beneficial.