I am seeing these big round structures in my RAW264.7 cell culture post-thawing. Does this mean the cells are unhappy? Any suggestions on what I can do about it?
They could be multinuclear fusions of your cells (synctia). I’m not sure how common that would be in your cell line-did they come from ATCC? Their tech service might help you.
Large, round structures can be indicative of cellular stress or changes in morphology due to suboptimal conditions. Cells can respond to various stressors by altering their shape. Also, Contaminants, such as mycoplasma can alter cell morphology and form structures that look different from the typical RAW264.7 cell. To address this, it is advisable to Check the conditions of your cell culture, including temperature, pH, nutrient availability, and any potential contamination. Ensure that your cell culture medium is appropriate for RAW264.7 cells. Consider performing viability assays to assess cell health
Susan C Hubchak they are from ATCC. Thanks for your response, I will reach out to ATCC tech service.
Ellen A G Chernoff I am using RPMI + 10% FBS (heat inactivated)
Gajendra Singh thanks for your advice
The RPMI medium is acceptabl,though DMEM is most common for this cell line. The morphology you see: multinucleated giant cells or osteoclasts with spread macrophage-like cells does not usually occur spontaneously, but requires stimulation with RANKL. Were these cells a high passage number that showed some differentiation before freezing? These cells do show genetic drift with high passage number.
Looks like "activated macrophages", ( may be "contaminations" or "dead cells" ).
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