I am culturing THP1 cells but I find most of the cells are getting dead and the cells which stay alive get attached to the plate even though it's not treated with PMA. Could anyone suggest a better solution for these? The media composition used is RPMI 1640 + 10% FBS + 1% antibiotics+ 50mM b-mercaptoethanol. Thank you.
Hi Kundan Solanki
50 mM beta-mercaptoethanol (BME) may be too high - I recommend using a final concentration of 0.5 mM BME instead.
Dear Kundan Solanki
I would like to share some tips on culturing THP-1 cells.
THP-1 cells are sensitive to cell density. You can passage cells between 3 x 10^5 and 7 x 10^5 cells/ml. Do not centrifuge THP-1 cells between passages. These cells will grow better in conditioned media. So, you leave at least 2ml of old media and add remaining fresh media to the culture.
Also, as mentioned by Jack Hopkins , the final concentration of beta-mercaptoethanol in culture should be 0.05mM.
Hope this will help you!
Good Luck.
Hi! We have a similar experience in the lab culturing THP-1 cells (cells were dying in RPMI). However, we have overcome with following media: RPMI 1640 Medium (ATCC modification) (cat. No: A1049101), 10% FBS, 1% antibiotics, 0.05mM b-mercaptoethanol. Cells are doing just fine now.
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