I am in the process of learning how to culture MSCs that I have flushed from the bone marrow of mice femurs. So far, the process runs smoothly until I attempt to trypsinize the cells, I attempt to do so 48 hours post initial culture, I have ensured the trypsin is at 37 degrees Celsius, and I return the culture to the incubator for the 3-5 minutes before neutralizing the trypsin. However, the majority of my MSC culture remains attached to the culture plate, with only a few cells lifting. Has anyone else had similar issues or some advice for this? I have also tried using cold PBS at 4 degrees Celsius, which had similar results. I have also used a cell scraper, but I'm not a massive fan of it due to the damage it can cause to the cells. Thanks in advance for any tips :)
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