Culture and identification of microvascular endothelial cells from human endometriosis

Abstract

To establish the methods of isolating and culturing human ovarian endometriosis-derived microvascular endothelial cells (OEMEC). The tissues of human endometriotic cyst of ovary were finely minced with scissors, then digested by collagenase I, II and trypsin-ethylene diamine tetraacetic acid (EDTA). The cells were purified by using centrifugation of 2000 r/min speed. OEMEC were identified by light microscope and transmission electron microscope observing CD(34), FVIII-Rag and Weibel-Palade in microvascular endothelial cells. The OEMEC grew as confluent monolayer like cobblestones under light microscope. CD(34) and FVIII-Rag were expressed strongly, and the percentages of CD(34) and FVIII-Rag positive cells were 91.4% and 92.5%. Weibel-Palade bodies could be observed under transmission electron microscope. The time of cell doubling proliferation was 4.5 days. The established system of isolating OEMEC would provide lab base for studying the mechanisms of angiogenesis in endometriosis lesions.