I recently purchased cryopreserved SW-982 cells. I thawed them according to the manufacturer’s protocol and cultured them in RPMI medium supplemented with 10% FBS, 1% antibiotics, and 1% glutamine (I’m unable to use L15 medium as my incubator is set to 5% CO2). From the start, the cells have been growing in suspension, forming spheres, with very few adhering to the flask surface.
I contacted the company, but they were unable to identify the cause of the issue.
Has anyone encountered this before? Why are they growing in this way, and what can I do to make them grow forming a monolayer?
Shayan Singh Parihar
- Thawing and Recovery Phase: Cells are often stressed after thawing and may take some time to fully recover and adhere. Did you change the medium 24 hours post thawing to remove any residual DMSO?. Try to increase the FBS concentration to 20% and observe for adherence.
- Confluency and Seeding Density: If the seeding density is too low, cells may struggle to adhere. Try increasing the seeding density slightly. After thawing, it’s often recommended to start with a higher seeding density to promote recovery and adherence.
- RPMI Medium: RPMI medium should be compatible with SW-982, but you might also consider testing other media like DMEM. While L15 medium is commonly used for these cells without CO2, you could try to optimize RPMI, and adjusting the CO2 concentration might also help (might not be possible in your case).
- Cell Quality: Might be that the cells were not handled correctly during transport and maybe damaged. Ask for a replacement from the manufacturer if this is the case. Do some troubleshooting, record the observations and submit it as proof to the supplier asking for replacment.
- Mechanical Detachment: Try gentle pipetting during media changes to see if this can break up the cell spheres and promote more even distribution and adhesion.
- Matrix or Surface Coating: Sometimes, certain cell lines benefit from a coated surface (e.g., collagen, poly-L-lysine, or fibronectin) to help with adhesion, especially after thawing. SW-982 cells don’t typically require it, but it may help in cases of attachment issues.
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