52 Decellularization of goat uterus as a promising 3-dimensional homing matrix of biological scaffold: A pilot study

Abstract

Female infertility or absence of a functional uterus has a prevalence of 3 to 5% in the general human population. Nowadays most patients diagnosed with absolute uterine factor infertility remain untreatable. Moreover, the world population is aging rapidly, and as people live longer, uterus frailty increases the risk of developing age-related diseases. Whole-organ decellularization and recellularization, a novel technique within the field of regenerative medicine, could eventually provide another solution compared with the reality of gestational surrogacy. Extracellular matrix to enable recellularization has been described using different animal models, where decellularization has been performed of whole uteri. The aim of the present study was to offer a new perspective with a goat uterus as a large whole reproductive organ model. For this purpose, uteri were collected from 6 goats after slaughter and attached to the perfusion circuit. Decellularization was achieved by 2 methods of whole-uterus perfusion, namely with Triton-X100 and dimethyl sulfoxide (group 1; n = 3), or with sodium deoxycholate (group 2; n = 3), both compared with a control group (n = 1). Morphological, immunofluorescence, real-time quantitative PCR analysis, coupled with vascular corrosion cast preparation, were used to assess protocol efficiency. Statistical analysis was carried out to identify significant differences between the 2 groups, using ANOVA and, when required, a post-hoc test (Bonferroni method). Morphological analysis showed different opacity between protocols. In both group 1 and 2, the removal of cells and cellular debris was confirmed with DNA quantification. Immunofluorescence confirmed the presence of the major extracellular matrix components, with collagen type I being the most abundant. Vascular tree network cast of the decellularized organ was successfully perfused in all the uteri. In conclusion, this preliminary study gives a new perspective in whole uterus bioengineering, utilising goats as a biological model to increase the knowledge in veterinary medicine as well as toward the in vitro and in vivo research of regenerative medicine in the field of reproduction.

Research was funded by the Carraresi Foundation.