Culturing cancer cells in a laboratory is a critical tool in cancer research. Scientists employ the method to study tumor biology, test drug responses, explore genetic variations, and more. Many methods of cell culture exist, one being primary culture. This method develops cultures from a sample from a living organism. In the case of cancer, it involves culturing cancer cells from a patient’s biopsied tumor. This guide outlines the basic steps to obtain a primary culture of cancer cells.
The Tools Involved
These are the main tools involved in primary culture development:
- petri dishes
- flasks
- pipettes
- culture media (substrate)
- enzymes like collagenase or trypsin
- antibiotics
- a biosafety cabinet
For the culture media, keep in mind that it must be tailored to the specific requirements of the cancer cell growth. For instance, collagen-coated culture dishes can bridge cells and substrate, potentially encouraging fast, prolific growth of the cancer cell culture.
Collecting Tissue Samples
Getting a cancerous tissue sample is typically done via biopsy or surgical excision of a tumor. The collected sample must move immediately to a suitable buffer or transport medium to preserve cellular integrity.
Disaggregating the Tissue
Disaggregation of the tissue will provide you with individual cells for culture. These are the basic steps:
Mechanical Dissociation
Use fine scissors or scalpels to cut the tissue into small fragments. This initial step reduces the size of the sample and prepares it for enzyme treatment.
Enzymatic Digestion
Incubate the tissue fragments in a solution containing enzymes like collagenase or trypsin. These enzymes break down the extracellular matrix, releasing individual cells. Monitor the digestion process closely and stop once the tissue has visibly dissociated.
Filtration
Pass the cell suspension through a sterile filter with an appropriate pore size to separate single cells from debris.
Culturing the Cells
After disaggregation, wash the cells with a fresh culture medium to remove enzymatic residues. Resuspend the cells in a complete culture medium that matches the nutritional and environmental needs of the cancer cell type. This may include fetal bovine serum (FBS), growth factors, and other supplements.
Seed the cell suspension into culture flasks or dishes at an appropriate density. Place the flasks in an incubator set to standard conditions to promote optimal growth.
Monitoring and Maintenance
Monitor the culture for cell growth, morphology changes, and contamination. Replace the culture medium every two to three days to replenish nutrients and remove waste. Maintain aseptic techniques throughout every step to preserve the integrity of the culture.
Obtaining a primary culture of cancer cells is an invaluable technique that supports much of modern cancer research. By following these steps, researchers can establish viable and reproducible cancer cell cultures to advance their studies.