Establishment and characterization of a novel human stromal cell line of peritoneal endometriosis

Endometriosis is a female gynecological disorder defined by the presence of endometrial tissue outside the uterus cavity. This complex, estrogen-dependent and inflammatory disease affects around 190 million women worldwide. Despite the high prevalence and costs of endometriosis, research of this disease is still underfunded and problems of unknown etiology, delayed diagnosis and lack of appropriate treatment remain unsolved. To improve understanding of endometriosis pathophysiology and discover novel diagnostic and treatment options for this debilitating disease, new experimental models of endometriosis are needed. The typical endometriotic lesion is histologically defined with the presence of epithelial glands and endometrial-like stroma. Since primary endometriotic cells have a limited lifespan, several immortalized cell lines have been created so far. However, currently available endometriotic cell lines are predominantly epithelial deriving from ovarian endometriomas. The present project proposal is focused on the establishment and characterization of a novel stromal cell line of peritoneal endometriosis. To achieve this, isolation and cell culturing of endometriotic primary cells will be optimized. Upon isolation, the purity of primary cell cultures will be determined using cell type-specific markers. The human telomerase reverse transcriptase (hTERT) plasmid will be applied for the creation of immortalized stromal cell line. The following characteristics of both primary cells and immortalized cell line will be assessed: viability, population doubling, cell migration, absence of cell senescence, absence of mycoplasma in cell culture medium, expression of morphological markers and estrogen-associated genes. Additionally, genomic stability using karyotype analysis and genetic authenticity using STR analysis of immortalized cell line will be determined. By establishing a well-characterized cell line, we expect to obtain a powerful experimental tool for studying endometriosis at the cellular level. This novel in vitro model of peritoneal endometriosis can be potentially valuable for drug screening or biomarker discovery.