I am culturing by 1640 basis medium (HEPES added, Glu added) + 10% FBS + 1% PS +0.1% β-ME, which taught by my colleague. Somehow, there is no one to freeze the THP-1 and I have searched many different formula medium for freezing THP-1.
Do I need to calculate the cell density before freezing them?
What is the optimal formula medium for freezing THP-1 cell? Can I make use of the freezing medium included 60%1640+30%FBS+10%DMSO? Or should I decrease the 10%DMSO to 5%DMSO? Did the 1640 medium is the basis one, without FBS and PS?
I've frozen THP1 in 40% RPMI 1640, 50% FBS and 10% DMSO with no problems. You want to to decrease the temp slowly, ie 1C per min or so, thus you should use either a glycol freezing container or insulate the vials with paper towels (ie roll the vial up 'n tape it tight). Overnight in -80, then liq N2. Good luck.
I have successfully frozen aliquots of 3x10^6 THP1 cells in FCS, 10% DMSO. Definitely use some sort of slow freeze system (eg a chryobox.) before moving them to liquid nitrogen.
Hello Miao, i have been working with THP 1 cells for more than two years. We culture them RPMI 1640 + 10% FCS. The cell density while giving a passage is generally kept as 3-4 million cells in a T 75 flask. We prepare freeze downs generally at P5 or P6 when the cells are quite stable , healthy and not experiencing any kind of stress. The freeze downs are prepared in FCS with 10 % DMSO. The cell density is kept around 3 million in 500 ul. After adding the freezing medium , the cryovials are immediately wrapped in cotton and transferred to -20 C, after one hour we transfer them to cryobox in -80 C. Up till today we have not encountered any kind of contamination or health issues of cellls in this protocol.
For further assistance you can check out ATCC guidelines.
Wish you luck !!
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