I am separating ovarian cancer cells from primary cell tissue and I do not have a FACS machine or any magnetic bead. Is there any suggestions to separate the fibroblast and mesenchymal cells from the ovarian cancer cells.
Hi Givon,
Immunopanning would be an option, but I'm not sure how expensive it can be.
Otherwise, shouldn't the cancer cell quickly overgrow fibroblast and mesenchymal cells? You might just wait for you cultures to be enriched.
Other option could be sub-cloning and isolating clones from individual cancer cells.
Cheers,
Lech
Givon - good morning. My first question is about the source of your tissue. Is this animal model material, or are you working with primary material from patients? If the latter, are you satisfied with your strategy for producing a more-or-less single cell suspension, or do you think you still have islands of cells and fibrosis that may make sorting difficult?
My second question is why magnetic beads aren't an option. Is it the lack of an appropriate antibody, or is there another reason?
If you do have an antibody with sufficient specificity for the cells you're looking for, microbubble separation would be another approach. We're working on this technology currently and have a 'universal' streptavidin-coated low-density microbubble that can buoyantly sort cells from complex suspensions. They've been used in positive selection (e.g., capture the cancer cell) and depletion (e.g., capture the fibroblasts and mesenchymal cells), so might be an additional option.
I'm happy to discuss the technology further. If it would be helpful, feel free to message me or reach out via the website.
Best, John
www.akadeum.com
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