Phenotypic and Functional Characterisation of Primary Endometriotic Stromal Cells for In Vitro Model Development

This study aimed to phenotypically and functionally characterise primary endometriotic stromal cells to support in vitro model development and the future establishment of an immortalised endometriotic cell line. Lesion tissues from 18 patients (ovarian, peritoneal, and rectovaginal septum lesions) were enzymatically digested using two protocols, then cultured and screened for mycoplasma, with assessments of viability, proliferation/population doubling time, migration (wound-healing imaging), gene expression (qPCR for oestrogen-related genes), immunocytochemistry markers (vimentin, cytokeratin, PAX2), 3D spheroid formation, and hormonal responses to oestradiol (E2) and medroxyprogesterone acetate (MPA). Cells maintained >90% viability through passage 5, showed migration kinetics largely unaffected by digestion protocol or menstrual cycle phase, but exhibited inter-individual variability, and had disease-associated features including downregulation of ESR1 and reduced ESR1/ESR2 ratio versus normal endometrium, stromal marker vimentin positivity with cytokeratin negativity, strong PAX2 expression, spheroid formation with variable morphology, and heterogeneous E2/MPA responses including reduced E2-induced growth linked to lower ESR1 and variable MPA effects consistent with progesterone resistance. The paper excluded four patient-derived cultures that failed to proliferate after isolation. This paper is centrally about endometriosis — it develops and validates primary endometriotic stromal cell cultures as an in vitro model, including their phenotypic markers and functional hormonal responses.

Read from the paper's body, not the abstract. Not a substitute for reading the paper. No clinical advice. How this works