Poster Presentation Hunter Cell Biology Meeting 2022

Human foetal female reproductive tract organoids to understand Müllerian duct anomalies (#89)

Varshini Devarapalli Venkata 1 , Muhammad F.B Jamaluddin 1 , Pradeep S Tanwar 1
  1. Global Centre for Gynaecological Diseases, The University of Newcastle, Newcastle, NSW, Australia

Background

Müllerian Ducts (MD) are paired tubular structures that give rise to the uterus, fallopian tubes, cervix, and upper portion of the vagina during embryonic development. Müllerian Duct Anomalies (MDAs) are developmental disorders of the female reproductive tract with a prevalence of 5% in fertile women and up to 6-15% in patients with infertility worldwide. Genome sequencing of patients combined with animal models has revealed the role of WNT signalling in MD development and differentiation, which is altered in patients with MDAs. However, key differences in mouse and human MD development limit the translation of these studies to patients. Therefore, human models are required to better understand the pathogenesis of MDAs and guide the development of personalized regenerative treatment for these patients.

Aim

To establish human foetal female reproductive organoids as a unique platform to investigate human female reproductive development and diseases.

Methods

In this study, we performed a comparative histopathology analysis to understand fundamental differences between human foetal and adult female reproductive tract epithelium. We developed human foetal female reproductive tract organoids from primitive fallopian tubes and uteri. Next, we compared the foetal organoids with their adult counterparts using histopathology analysis and proteomic analysis. To assess the regenerative potential of foetal organoids, we performed transplantation and regeneration of decellularized adult tissue scaffolds by foetal organoids. Finally, the transplanted tissues were treated with WNT inhibitors to define the role of WNT signalling in human female reproductive tract development and diseases.

Results

Histopathological and proteomic analysis revealed significant differences in protein expression and signalling pathways between human foetal and adult reproductive tract tissues. We showed the successful establishment of culture conditions for foetal organoid growth and long-term maintenance. Our culture conditions preserve the in vivo tissue characteristics and promote the in vitro differentiation of immature foetal epithelium. The organoid-scaffold model provided evidence that the foetal organoids comprising the immature epithelial progenitors represent a transplantable source of cells and have the capacity to regenerate the adult organs. Additionally, we highlighted the requirement of WNT signalling in governing the self-renewal and differentiation of the foetal epithelium.

Conclusion

To conclude, we have developed precious resources of human foetal female reproductive organoids to understand the basis of human female reproductive development and associated disorders. In the future, the organoid-scaffold model can be used to provide better tools to improve the current regenerative treatments available for patients with MDAs.

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