Abstract
Endometrial organoids (EMO) are an important tool for gynecological research but have been limited by generation from (1) invasively acquired tissues and thus advanced disease states and (2) from women who are not taking hormones, thus excluding 50% of the female reproductive-aged population. We sought to overcome these limitations by generating organoids from (1) menstrual fluid (MF; MFO) using a method that enables the concurrent isolation of menstrual fluid supernatant, stromal cells, and leukocytes and (2) from biopsies and hysterectomy samples from women taking hormonal medication (EMO-H). MF was collected in a menstrual cup for 4–6 h on day 2 of menstruation. Biopsies and hysterectomies were obtained during laparoscopic surgery. Organoids were generated from all sample types, with MFO and EMO-H showing similar cell proliferation rates, proportion and localization of the endometrial basalis epithelial marker, Stage Specific Embryonic Antigen-1 (SSEA-1), and gene expression profiles. Organoids from different disease states showed the moderate clustering of epithelial secretory and androgen receptor signaling genes. Thus, MFO and EMO-H are novel organoids that share similar features to EMO but with the advantage of (1) MFO being obtained non-invasively and (2) EMO-H being obtained from 50% of the women who are not currently being studied through standard methods. Thus, MFO and EMO-H are likely to prove to be invaluable tools for gynecological research, enabling the population-wide assessment of endometrial health and personalized medicine.
Highlights
The diagnosis and study of gynecological disease currently utilizes endometrial biopsies that are often acquired under general anesthesia
We aimed to compare the endometrial organoids that were derived from endometrial biopsies or hysterectomies (EMO) to those derived from menstrual fluid (MFO) and from endometrial tissue from women taking hormonal medication (EMO-H)
The amount of glandular material that was available for organoid generation was often lower in the menstrual fluid (MFO) and endometrium from the women who were on hormones (EMO-H) compared to the endometrial biopsies from the women who were not on hormones
Summary
The diagnosis and study of gynecological disease currently utilizes endometrial biopsies that are often acquired under general anesthesia. Endometrial biopsies do not provide information about the function of endometrial cells, which are important for fertility and are implicated in diseases such as endometriosis, adenomyosis, Asherman’s syndrome, and infertility. While colony forming assays allowed the identification of endometrial epithelial stem/progenitor cells [2,3,4], the study of the endometrial epithelium was 4.0/). Curtailed by an inability to maintain those cells in long-term culture. The generation of organoids from endometrial tissue [5,6,7] has enabled the long-term culture of endometrial epithelial cells, aiding our understanding of endometriosis [6] and fertility [8]
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