Method for Obtaining Primary Ovarian Cancer Cells From Solid Specimens

Establishment of Epithelial Cell Culture from Ovarian Cancer Tissues: A Method Comparison Study

“…The previous studies of epithelial ovarian cancer cell culture were performed using different methods of cell dissociation and different media. Studies conducted by Pribyl et al,(2014) and Kar et al, (2017) utilized Dispase and DMEM:F12 medium to establish the ovarian cancer primary cell culture. In another study conducted by Shepherd et a., (2007) the ovarian tissues were processed into cell suspension using Dispase and precipitated using Percoll, and then the cells were cultured in MCDB105:M199 medium.…”

“…Dispase is a natural protease that is isolated from a bacteria, Bacillus polymyxa (Stenn et al, 1989). This enzyme is widely used for dissociating tissues for cell culture application including epidermis (Stenn et al, 1989), pancreas (Ono et al, 1977), spleen (Fujiyama et al, 2018), brain cells (Volovitz et al, 2016;Zhu et al, 2012), stem cells (Jager et al, 2019), and cancerous tissues such as the mammary (Nishikata et al, 2013), lung (Kim et al, 2013), and ovary (Pribyl et al, 2014;Shepherd et al, 2007). Dispase specifically breaks down fibronectin, which typically assembles integrins that bind cells in the tissues, and type-IV collagen, which constructs the basal lamina of tissues, indicating that this enzyme has higher specificity compared to trypsin (Stenn et al, 1989;Zhu et al, 2012).…”

“…In searching the most suitable models for ovarian cancer, we investigated the feasibility of developing primary cell cultures directly from patients' tissues, expecting that the cultures will highly mirror the real molecular events of the cancer. Previous studies have established (O'Donnell et al, 2014)methods to obtain primary cell culture from ovarian cancer tissues by utilizing combination of enzymes, such as Dispase and Trypsin, and culture media, such as DMEM:F12 and McCoy's 5A (Kar et al, 2017;Pribyl et al, 2014;Shepherd et al, 2007;Thériault et al, 2013). However, the epithelial cells of many tissues, including ovarian cancer, are often difficult to be cultured and oftentimes became outgrown by the fast-growing fibroblast (Chen et al, 1993;Hongpaisan, 2000).…”

“…However, the epithelial cells of many tissues, including ovarian cancer, are often difficult to be cultured and oftentimes became outgrown by the fast-growing fibroblast (Chen et al, 1993;Hongpaisan, 2000). This study aims to develop the most efficient protocol to raise epithelial cells from serous ovarian cancer tissue by comparing several combinations of cancer tissue site (primary or metastasis), different cell suspension methods, the use of percoll, and the type of culture media, that were demonstrated separately in the previous studies (Kar et al, 2017;Pribyl et al, 2014;Shepherd et al, 2007;Thériault et al, 2013).…”

“…All cancer tissues were processed to make cell suspension and later to be derived into primary cell cultures using several combinations of methods , performed by previous studies (Kar et al, 2007;Pribyl et al, 2014;Thériault et al, 2013). The primary and metastatic tissues were experimented according to Figure 1a (for primary site) and figure 1b (for metastatic site).…”