atively few in vitro and in vivo models available for studying the com plex disease biology, pathophysiology, and for use in the preclinical development of novel therapies. One widely used in vitro model comprises cell lines of epithelial and stromal origin harvested from peritoneal endometriosis lesions. Co injection of these cells into mice results in peritoneal lesions that recreate histological features of human endometriosis in vivo. This particular model has been used to study various as pects of endometriosis cell biology including hormone sig naling, cell cell adhesions, as well as to conduct candidate gene studies. However, this model was established from a peritoneal lesion, and there is a real need for add itional models that mimic other subtypes of this disease in order to better understand the pathophysiology of endo metriosis subtypes, and for the development of new treat ment strategies.

A major limitation of existing in vitro models of endo metriosis is that they have been established by culturing endometriosis epithelial cells as monolayers on tis sue culture plastics selleck chemicals cultures. In vivo, EECs exist within a dynamic three dimensional microenvironment and constantly interact with a stroma containing immune cells, fibroblasts, vasculature and a heterogeneous network of extracellular matrix. Endo metriosis cells in vivo also form cell cell interactions through the entire cell surface. By contrast, cells in 2D can only interact along a small proportion of the plasma mem brane.

Several studies have now reported on improvements in in vitro modeling of several diseases when target cells are cultured as 3D models, cultured cells maintained in 3D re semble the tissues selleck chemical of origin more closely than the same cells cultured in 2D. However, to our knowledge, there are no studies reporting in vitro spheroid models of endometriosis. Such models could be particularly useful for developing novel therapies for this disease and for studying the links between endometriosis and ovarian cancer. To better model the biology of ovarian surface endomet riosis, we have established and characterized a novel endo metriosis epithelial cell line, EEC16, from a 34 year old patient diagnosed with ovarian endometriosis. EEC16 and a second endometriosis cell line from a pa tient with peritoneal endometriosis were established as in vitro 3D cell culture models and the morphological and molecular features evaluated.

EECs grown as 3D cultures mimic endometriosis lesions in vivo more closely than 2D cultured counterparts, suggesting that these models are ro bust representations of human endometriosis for future use in understanding the etiology of endometriosis and identi fying novel therapeutic targets for the disease. Methods Primary tissue collection Endometriosis cells we