As the cell density usually higher than culture medium, I just wonder how can suspending cell lines won't attach the flask bottom?
Hi, they are not adherent cells. Once they differentiate in macrophages, they become adherent. You can differentiate them with PMA. If they are not adherent, it means that your culture is probably ok (some contaminations make them adherent). You can check their viability and structure under the microscope to confirm that they are healthy.
Thanks so much for your answers. But what I concerned is not how to stimulate THP-1 adhere to substrates. I just wonder why THP-1 can suspending in the culture medium when their density higher than culture medium. Thanks in advance!
Hey, if I have correctly understood your question, it is about stopping the monocytes from differentiating into adherent macrophages. If that is the case then take care of the following points and you will be able to maintain them.
1. Keep the cell density lower than 106 cells/mL media always. Distribute the cells into two as soon as they reach close to this density.
2. Do not forget to add beta-mercaptoethanol to your cells as they will help them stay away from stress and keep them happy in their original morphology.
3. RPMI-1640 medium has always worked the best for THP-1 in my lab.
4. Do not add excess antibiotics to your culture.
After doing all of these also, I am afraid if you can use your THP1 flask for more than 15 passages. They tend to start changing their morphology sooner than other cell lines.
Please write if you wish to know the concentration of culture media components or company.
Best luck!
Urja Verma Thanks so much for your answer! Maybe I didn't illustrate my question clearly. Actually, I just curious why THP-1 cells (not in macrophages) can suspend in culture medium (say RPMI1640+FBS). As I know, the THP-1 cell density is higher than culture medium. It should be contact (not adhere) on the substrate of culture flask due to the gravity. Thanks again for your attention.
Hi, oh yes, interestingly the non-adherent cells settle less in the culture vessels. Especially if you talk about the low-density cells. One of the low-density cells is monocyte. Their specific gravity is below 1.1 g/mL. The density of culture media, if lesser than that of the cells, they may not settle on the bottom. Therefore, measuring the approximate density of your media may help you to feed your curiosity about why your cells are not settling down!
Cheers!
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