Isolation and Characterization of Primary Tumor Cells
The lack of effective “in vitro” models to predict success of pharmacological therapy for patients with variety of cancer cell types demands the development of better experimental models to understand the pathophysiology of the disease and evaluate drug sensitivity in vitro. Cell lines are well characterized homogenous population that gives consistent and reproducible result from experiment to experiment. Many of these cell lines were established over 20 years ago. Many agents with impressive activity in cell lines show little clinical effect thereby making the data insufficient to predict efficacy in the clinic. Therefore this study on primary cancer cell lines will help improve the accuracy of the results during drug development. With this in view, 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 and most of 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. Therefore these studies with the primary cell lines will help in obtaining biological responses mimics the tumor/cancer micro environment more accurately
Collaborators: Dr. Lakshmi S, Regional Cancer Center (Trivandrum)
