In Vitro Culture of Primary Cells from Cancer Patients- Screening for Potent MMP-9 Inhibitors
Publication Type:Conference Paper
Source:The XXXIX All India Cell Biology Conference (2015)
Breast cancer is the most prevalent form of cancer among women worldwide. Established cancer cell lines generated over years have been used for studying the biology of breast cancer. These cell lines which have been passaged innumerable times creates genetic drift which makes the observations biologically less relevant. In vitro culture of primary cells from fresh surgical specimens provides a more accurate means for understanding the behavior of cancer cells and the adjacent normal cells. We have developed a simple and rapid method for isolating primary cultures from breast tumor and normal cells. Two different methods were employed for isolating primary cultures- one involving enzymatic digestion and the other using explant culture which is independent of enzymatic treatment and feeder cells. The primary cells obtained were further characterized by immunocytochemistry staining for Vimentin and Cytokeratin 18. The cells stained positive for Vimentin and negative for Cytokeratin 18 which indicated that they are of mesenchymal origin. Since gelatinases play a prominent role in promoting breast cancer metastasis, the primary cells were assessed for gelatinase activity. Our observations clearly show that the cancer cells had up-regulated gelatinases activity compared to the adjacent normal cells. These cells when treated with several natural products showed a dose-dependent inhibition of gelatinase activity. The results obtained were consistent in all the primary cell lines generated from different patient samples. These studies with the primary cell lines will therefore help in obtaining biological responses that more accurately mimics the tumor/cancer micro environment.
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