Principles of scaffold generation for bioengineering of the ovary and uterus: a study focusing on decellularization

Abstract

Introduction: Cancer therapy often result in fertility problems due to inflicted injury to the reproductive organs. Since most women survive cancer, fertility preservation has become an important consideration during cancer therapy. However, options for young women with blood-related cancers are missing. Papers I-II describe the development and characterization of mouse ovarian scaffolds derived from ovarian extracellular ma-trix (ECM). Such scaffolds may be used for future ovarian bioengineering applications as a supporting matrix for the expansion of immature follicles isolated from young can-cer patients to preserve their fertility. Paper III-IV use the rat model to analyse similar scaffolds for uterus bioengineering applications and evaluate if these scaffolds are im-munologically inert after engraftment. Methods: Three decellularisation protocols based on sodium dodecyl sulfate (SDS) and sodium deoxycholate (SDC) were developed for mouse ovary scaffold production (Pa-per I). Scaffolds were then characterised using histology and quantitative analysis for ECM components. Recellularisation was tested using mesenchymal stem cells (Paper II). Previously established uterus scaffolds were grafted to syngeneic (Paper III) or al-logeneic rats (Paper IV) to investigate if the decellularisation process generated any detrimental damaged associated molecular products (DAMPs; Paper III) and if the al-logeneic recipient’s immune system remained stable after scaffold engraftment (Paper IV). Immunohistochemistry and gene expression analysis with digital droplet PCR was used quantify infiltrating immune cells and expression of proinflammatory signals. Results and conclusions: Paper I developed three novel mouse ovarian scaffolds. The SDS and the SDC protocols were found promising, whereas a protocol based on both detergents were found too aggressive on the ECM. Paper III showed that a mild, yet effective decellularisation protocol generated less amounts of DAMPs, and that this scaffold type also remained the most inert to the recipient’s immune system in an al-logeneic setting (Paper IV).