Fabrication of Endometrial Cell Sheet for Fertility Resumption by a Transgenic Mouse Model

Human endometrium is a dynamically remodeling tissue which undergoes more than 400 cycles of regeneration, differentiation, and shedding throughout a woman’s reproductive life. Trauma to the endometrium, including the basalis and underlying myometrium may result from postpartum curettage, spontaneous miscarriage, termination of pregnancy, or endometrial ablation procedures. Deep trauma involving the basalis and underlying myometrium may explain the failure of the endometrial functional layer to regenerate in Asherman syndrome and intrauterine adhesions (IUA). The failure to construct a functional endometrium in patients with Asherman syndrome is one of the main unresolved clinical issues in reproductive medicine. Although endometrium reconstruction shows an importance, so far there is no effective way to restore a functional uterus in clinical till now. A couple of years ago, we proposed a new way to engineer cell sheet (multi-layered cells with extracellular matrix). Briefly, we used a transparent PVDF membrane grafted disulfide bond-containing amino acid and biopolymer in order, and the surface modified transparent PVDF membrane can be cleaved to achieve a detaching system between disulfide bonds via a reduction by amino acidic reductant. The in vivo bioluminescence (BLI) technology allows non-invasive longitudinal imaging of markers of gene expression in vivo by luciferase-catalyzed reactions. Due to its high sensitivity, it is suitable for integration into research in transplantation and regenerative medicine. It can provide the real-time information of tissue viability. Moreover, no control mice are needed, so animal requirements are lessened because continual time measurements can be made of the same whole, living animal. So, this grant aimed to employ in vivo BLI in living transgenic mice as an animal model to reconstruct endometrial tissues in vitro and ectopically regenerate endometrium-like tissue in vivo using cell sheet engineering. This study will then designed to determine endometrium-specific functions by analyzing histology and mRNA expressions in endometrium-like tissue. The transgenic mice’s endometrium-like tissue will be implanted into a wild-type mice then to determine endometrium functions by BLI under in vivo imaging system (IVIS). Throughout the three-year grant of execution, we wish these obtained results will be useful for the next step on clinical application.