THP-1 cells culture - (Nov/20/2007 )

Hi,

Does any of you have some good rules about the conditions to culture THP-1 cells.

- Type of flask used (T75???)
- In which way? (vertical of horizontal like other cell types?)
- What is the best seeding concentration, which volume? Usually with adherent cells, I use about 20 ml of cell culture media in a T75 flask. Does this change with suspension cells?
- What is the density that we need to pass the cells and which interval is needed to change the medium?
- How many passage we can do to do not modify cell behavior? I know that transformed cells are "eternal", but it is the case with these cells?
- I saw somewhere that it is necessary to add 2-mercaptoethanol to a final dilution of 0.05 mM, why using this?
- How do you put cells in starvation? RPMI 1640 medium with 10% FBS, do we have to remove only the FBS or it is necessary to maintain a little amount of serum in the media?

Thank you so much for these precious informations.

Brassap

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I use normal cell culture flasks which I then incubate horizontal.

I splitt my cells by eye. Mostly 2-5 ml per 30 ml media. I splitt them about 40 times bevore I trash them. But sometimes they behave odd already at 30 passages.

I never added 2-mercaptoethanol to my media.

Starvation I use with 0.3 % of FCS.

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Thank you very much for these useful specifications.

Have a great day!

Brassap

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Dear Brassap


Another word of warning is to NOT freeze the cells ( THP-1) with DMSO, as this can differentiate the cells. Always use glycerol.

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Thank you very much for this important precision.

Have a great day!

Brassap

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