hello,
Is it possible to culture RAW264.7 and THP 1 cells without 10% serum ? If that can for how long do they live? Where can I find answers for these kind of questions? Can any one suggest a book or a web? or give me an answer?
thanks
With RAW264.7 cells, be careful about culturing for too long with little to no serum. The cells will become highly vacuolar and stellate, which indicates stress in this cell line. Their responsiveness to death stimuli will be increased. You can culture in low serum medium (1-2%), but you might have to change the medium more frequently.
RAW264.7 are deficient in certain proteins which can stimulate death under stress conditions (such as ASC/PYCARD), but THP-1 are not. This might lead to stress and death in THP-1 even sooner, though I have less (but some) experience with that cell line.
Both cells can be cultured for short period of time in serum depleted medium. However, when you look for some functions it is advisable to check for the effect of serum.
Yes, you can definitely culture both these cell lines without serum for 24 hours of experiments. I have done it for beta amyloid peptide treatments.
You can read my papers as well
Jordan , Senthil thank you for your help I just want to grow them for 3 days without serum do any one of you have experience with culturing these cell lines without serum? how long did they live? they can reach 3 days of growing ?
Zina, We have cultured RAW264.7 in 10% FBS in culture flasks. However for cytokine release assays, we have first cultured them for 24h in 10%FBS in culture plates. Subsequently, we have incubated these cells even in 0% FBS for 24 h to bring down cytokine levels to basal and then stimulate with inflammatory agent such as LPS/ConA to see enhanced secretion of cytokines. Cells look absolutely happy and with good morphology!
And yes, after serum-starvation, we have treated cells with LPS+anti-inflammatory agent for a duration of maximum of 48 h, so the total duration without FBS is 3 days; 72 h. Cells look perfectly fine!
For THP-1, if you use 5% FBS, they can survive up to 4 days (started cells density is around 1 million cells per ml)
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