ESA-SRB-AOTA 2019

Quantification and clonogenicity of stem/progenitor cells in menstrual fluid from healthy women (#117)

Caroline E Gargett 1 2 , Sophie Suke 1 , Caitlin Filby 1 3 , Jemma Evans 4
  1. Obstetrics and Gynaecology, Monash University, Melbourne, Victoria, Australia
  2. The Ritchie Centre, Hudson Institute of Medical Research, Melbourne, Victoria, Australia
  3. The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia
  4. Centre for Reproductive Health, Hudson Institute of Medical Research, Clayton, Victoria, Australia

Shedding and regeneration of the endometrium is critical for embryo implantation. Disordered shedding (endometriosis, adenomyosis) and/or regeneration (Asherman’s Syndrome) is associated with debilitating menstrual disorders. Endometrial stem/progenitor cells, likely responsible for endometrial regeneration, are SUSD2+ mesenchymal stem cells (eMSCs) and N-cadherin+ (NCAD) epithelial progenitors (eEPC). NCAD+ eEPC form gland-like structures in organoid cultures, self-renew and are located in the bases of endometrial glands. SUSD2+ cells are perivascular cells in the endometrium. SUSD2+ eMSC and NCAD+ eEPC are shed during menstruation in women with/without endometriosis (unpublished). Our aim was to determine if there are biological variations in endometrial stem/progenitor cell concentrations and clonogenicity in menstrual fluid between menstrual cycles in normal women and between women.

Menstrual fluid (day 2) was collected from healthy women (not on hormones; regular cycles) in a menstrual cup. Endometrial cells were dissociated with enzymes, leukocytes depleted by CD45 magnetic beads and red blood cells by hypotonic lysis. The %SUSD2+ and %NCAD+ cells were determined by flow cytometry and clonogenicity by colony forming assays (cell seeding, 50/cm2).

Menstrual fluid was collected from 3 women over 3 cycles. SUSD2+ cells were present in all menstrual fluids (1.0-7.1% of CD45- cells; median 5.4%). NCAD+ cells were detected in 5 of 7 samples (0-3.0% of CD45- cells; median 0.6%) with no variation between cycles or participants (p=0.4). The clonogenicity of CD45-endometrial cells (3.9% - 17.8%) was also similar across cycles and between women.

In this pilot study, there was a lower frequency of NCAD+ cells compared to SUSD2+ cells in menstrual fluid as expected from their location. Further samples are required to fully determine the variability of endometrial stem/progenitor cells shed between women and between cycles of individual women.  Quantification of endometrial stem/progenitor cells in menstrual blood may be a simple, non-invasive method for prognosis of fertility and endometrial disorders.