Development and characterization of human fetal female reproductive tract organoids to understand Müllerian duct anomalies

Abstract

Müllerian ducts are paired tubular structures that give rise to most of the female reproductive organs. Any abnormalities in the development and differentiation of these ducts lead to anatomical defects in the female reproductive tract organs categorized as Müllerian duct anomalies. Due to the limited access to fetal tissues, little is understood of human reproductive tract development and the associated anomalies. Although organoids represent a powerful model to decipher human development and disease, such organoids from fetal reproductive organs are not available. Here, we developed organoids from human fetal fallopian tubes and uteri and compared them with their adult counterparts. Our results demonstrate that human fetal reproductive tract epithelia do not express some of the typical markers of adult reproductive tract epithelia. Furthermore, fetal organoids are grossly, histologically, and proteomically different from adult organoids. While external supplementation of WNT ligands or activators in culture medium is an absolute requirement for the adult reproductive tract organoids, fetal organoids are able to grow in WNT-deficient conditions. We also developed decellularized tissue scaffolds from adult human fallopian tubes and uteri. Transplantation of fetal organoids onto these scaffolds led to the regeneration of the adult fallopian tube and uterine epithelia. Importantly, suppression of Wnt signaling, which is altered in patients with Müllerian duct anomalies, inhibits the regenerative ability of human fetal organoids and causes severe anatomical defects in the mouse reproductive tract. Thus, our fetal organoids represent an important platform to study the underlying basis of human female reproductive tract development and diseases.

Generation of Human Endometrial Assembloids with a Luminal Epithelium using Air–Liquid Interface Culture Methods

“…Other LE markers, including VTCN1 [ 19 ] and MSLN , [ 25 , 41 ] were also specially expressed in the putative LE subpopulations (Figure 6a,b ). Conversely, HEY1 , [ 19 , 42 ] SCGB2A2 [ 25 ] and FOXA2 [ 19 , 25 ] were highly expressed in putative GE subpopulations (Figure 6a,b ). The similar expression patterns of LE‐ and GE‐specific markers (Figure 6a,b ) suggested that the distribution of the LE‐ and GE‐like subpopulations in ALI‐EnAos was consistent with the in vivo endometrium, as defined by previous reports.…”

Harnessing three-dimensional (3D) cell culture models for pulmonary infections: State of the art and future directions

“…Van Riet et al cultured microfluidic lung chip model from alveolar type 2 cells isolated from resected lung tissues of emphysema patients who underwent surgery. AEC2 cells processed were cultured into organoids using alveolar culture medium and seeded on both, inserts and chips for comparison, and it was found that lung-chip could be a more reliable model than conventional cell culture inserts (van Riet et al 2022 ).…”

Proteomic and functional characterization of intra-tumor heterogeneity in human endometrial cancer

“…Organoids from freshly collected tissue biopsies from four patients (16 biopsies from four patients) were developed and maintained using a published protocol. 81 , 82 Briefly, fresh tumor biopsies were washed with PBS (PBS) containing 1% penicillin/streptomycin and amphotericin to remove excess blood. Tissue samples were cut into small pieces using a scalpel blade and incubated with accumax (Thermo Fisher Scientific) for 45 min at 37°C on a shaker.…”