Good day, I have been recommended to expand my suspension cells, specifically, the THP-1 cells by seeding 5 million cells in 100mL RPMI 1640 media and wait for around 1 week and a half until cell density reaches 1,000,000 cells/mL. I was wondering if this is the norm, and what other methods are they? Are THP-1 cells very slow dividing cells?
Also, the flask would be in an horizontal or vertical position?
Thank you.
That's the gist of it. It shouldn't take 10 days, the doubling time is 24-30 hours, so going from 5 million to 100 million should take 5-6 days.
That being said, the important thing is to start with a good density of cells, THP-1 are quite sensitive to cell density. You're at 0.5e5/mL (5 million / 100 mL). I would probably recommend double that 1e5/mL or even 2e5/mL.
Also make sure your RPMI 1640 medium has 2 mM L-glutamine, 10% FBS, and I also added 0.05 mM 2- mercaptoethanol, but it's not clear this even does anything. For thawing cells, you may even consider using 20% FBS initially.
Keep your flask horizontal, since this ensures the liquid is shallow and permits better oxygen transport due to better surface area to volume ratio. Just follow the guidelines for your flask as to the volumes.
Hello, Thp1 is relatively slow growing cells but you can accelerate its growth by adding 10% v/v of heat in activated bovine serum and and also by adding glutamine, incubate at 37c and the flask should be horizontal to give more space for cells and avoid cell overcrowding, when you are washing your cells try to use preheated Rpmi to avoid heat shock and then daily check the flask to see whether your cell are happy or no
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