Obtaining Primary Ovarian Cancer Cells from Solid Specimens: A Method to Culture Epithelial Ovarian Cancer Cells from Ovarian Tumor Specimens
Transcrição
Epithelial cells present on the outer surface of the ovary may undergo malignant transformation to cancerous cells that can grow into ovarian tumors.
To isolate these epithelial ovarian cancer cells or EOCs, begin by taking an ovarian tumor specimen and mince it into small pieces. Transfer the minced pieces into a tube containing a digestion buffer supplemented with the desired protease enzyme.
These proteases degrade the extracellular matrix proteins within the tissue, releasing EOC cells, erythrocytes, and a few dissociated fibroblasts into suspension.
Filter the cell slurry through a strainer to separate the suspended single cells from undissociated tissue pieces. Centrifuge the suspension to pelletize the EOC cells, erythrocytes, and fibroblasts. Discard the enzyme-containing supernatant.
Resuspend the cell pellet into an epithelial growth medium and transfer it into a culture dish. Incubate the culture to allow the EOC cells and fibroblasts to attach to the base of the dish while the erythrocytes float in the media.
Remove the suspended erythrocytes by refreshing the media. Over time, the media facilitates the selective growth of EOC cells over fibroblasts to form a monolayer of primary ovarian cancer cells.
