Bioengineered uterine tissue to support pregnancy in a rat model

摘要

Introduction: Absolute uterine factor infertility due to congenital/surgical absence of the uterus, or due to malformation or intrauterine adhesions affects about 1:500 women. Our group reported the first ever live birth after human uterus transplantation last year and this has now been followed by more births. Thus, this previously untreatable condition can now effectively be cured although with associated problems of risky donor surgery, organ shortage and side-effects of immunosuppression. The creation of a bioengineered uterus would circumvent these issues. In our animal-based research towards a human bioengineered uterus we initially use the rat model. In this study we have explored the creation of bioengineered uterine patches for tests in vivo as a step towards the creation of a complete bioengineered uterus, but also to test this patch-concept as a way of uterine-repair of a partially defect uterus. Methods: Decellularization was achieved by whole-uterus perfusion through the uterine arteries with bufferedon-buffered Triton-X100/DMSO (Group 1/2) or sodium deoxycholate (Group 3). Recellularization was achieved by injecting GFP labelled mesenchymal stem cells and uterus primary cells into the patches (5×10mm). Three days later one patch per uterus was transplanted into SD-rats to repair created uterus defects, and 6 weeks later the animals were mated. The experiments were terminated 16-20 days after mating to assess pregnancy results (number and sites of fetuses; immunohistochemistry; qPCR). Results and Discussion: Successful recellularization was confirmed by fluorescence and immunohistochemistry, but revealed limited cell distribution. Pregnancy and fetal development was normal in Group 1 and 2, with fetal development over patch areas. Group 3 showed a significant reduction in fetal numbers and no fetal development in the graft area. qPCR and immunohistochemistry revealed uterus-like tissue in the patches, with a remodulation by infiltrating host cells. Conclusion: The functionality of the reccellularized Triton-X100/DMSO-generated scaffolds was near normal and these protocols should be explored further towards the development of bioengineered uterine patches to treat partially defect uteri.