The Notch and Wnt pathways regulate stemness and differentiation in human fallopian tube organoids

Abstract

The epithelial lining of the fallopian tube is of critical importance for human reproduction and has been implicated as a site of origin of high-grade serous ovarian cancer. Here we report on the establishment of long-term, stable 3D organoid cultures from human fallopian tubes, indicative of the presence of adult stem cells. We show that single epithelial stem cells in vitro can give rise to differentiated organoids containing ciliated and secretory cells. Continuous growth and differentiation of organoids depend on both Wnt and Notch paracrine signalling. Microarray analysis reveals that inhibition of Notch signalling causes downregulation of stem cell-associated genes in parallel with decreased proliferation and increased numbers of ciliated cells and that organoids also respond to oestradiol and progesterone treatment in a physiological manner. Thus, our organoid model provides a much-needed basis for future investigations of signalling routes involved in health and disease of the fallopian tube.

Oviduct: roles in fertilization and early embryo development

“…Notch and WNT signals not only modulate the oviduct development but also mediate the differentiation of adult epithelial stem cells into other cells in Fallopian tubes (Kessler et al 2015). Inhibition of Notch signaling by a γ-secretase inhibitor, dibenzazepine, in the oviduct and the stem-like epithelia leads to a genetic signature of cell differentiation into ciliated epithelium.…”

Development of organoids from mouse and human endometrium showing endometrial epithelium physiology and long-term expandability

“…We dissociated mouse endometrium and cultured the glandular-type fragments (Fig. S1A) in conditions previously shown to enable organoid formation from a diversity of tissues (Barker et al, 2010;Karthaus et al, 2014;Kessler et al, 2015;Ren et al, 2014;Sato et al, 2009Sato et al, , 2011. Typically, dissociated tissue is embedded in Matrigel as an extracellular matrix scaffold, and cultured in a cocktail of growth and signaling factors, essentially containing WNT activators like WNT3A; R-spondin 1 (RSPO1), which is the ligand of leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5) and acts as a WNT signaling amplifier; epithelial-cell mitogens, such as epidermal growth factor (EGF) and fibroblast growth factor 10 (FGF10); inhibitors of bone morphogenetic protein (BMP) such as Noggin, which allow long-term expansion by preventing differentiation; and antagonists of the transforming growth factor β (TGFβ) pathway such as A83-01 Alk inhibitor, which maintain epithelial-cell character.…”

“…Given the well-known mitogenic effect of insulin on endometrial epithelium (Shiraga et al, 1997), our medium was further supplemented with insulin-transferrinselenium (ITS). As a source of WNT pathway activation, we used WNT3A-and RSPO1-conditioned media (CM), both added at 25% v/v (Kessler et al, 2015). The final culture medium (for full composition, see the supplementary Materials and Methods) is referred to as 25W/25R (indicating the presence of 25% WNT3A-CM and 25% RSPO1-CM).…”

Translational potential of human brain organoids

“…This landmark paper established a novel experimental paradigm for generating tissue organoids. Subsequent studies worldwide confirmed the generalizability of such approach to human stem cells, and produced organoids of a variety of organ types including colon,11 intestine,12 prostate,13, 14 fallopian tube,15 stomach,16 liver,17, 18 kidney,19 lung,20 and brain 8…”