I am culturing THP1 cells in RPMI 1640 (as advised by ATCC formulation, except 2ME). Unfortunately my cells are forming clumps a few hours after seeding (at 1* 10^5 cells/ml) before even going to 60% of confluence. I am using pen-strep in the media to prevent bacterial contamination. After they form clumps, they stop growing. How can I get over this problem? Thank you for your help.
How big are the clumps? THP-1 cells grow in clumps as when they divide the new cell attaches to the one it split from. With my THP-1 gentle pipetting up and down will break the clumps but the cells remain viable. If you split the culture often you should have small clumps 2-4 cells, if you leave them for a few days the clumps will grow larger as the cells replicate.
Hi,
It is possible that adding 2-mercaptoethanol, as advised by ATCC, can help you with this problem. Did you check the viability of your clump-forming cells?
Thank you for the suggestion. I have tried with 2 mercaptoethanol. But there was no improvement in respect to cell clumps rather it caused more debris. Cells were fine in Trypan Blue test and they were responding to PMA induction. But i am not getting enough cells to perform full length experiment. May be clumping formation is preventing cells to grow. How to get rid of this problem? plz help!
How big are the clumps? THP-1 cells grow in clumps as when they divide the new cell attaches to the one it split from. With my THP-1 gentle pipetting up and down will break the clumps but the cells remain viable. If you split the culture often you should have small clumps 2-4 cells, if you leave them for a few days the clumps will grow larger as the cells replicate.
Thank you for suggestions! My THP1 is forming bigger clumps (about 30-40 cells together) after one day of seeding. There are less numbers of single cells and number of clumps increases over time. When i count the cells after 2 days of culture there is not even 2 fold change in number. What could be the possible reason? plz help..
Same problem here! Here is a Onedrive link to a photo of how my cells look:
http://1drv.ms/1efCtab
My cells keep growing normally and the population doubling time is about 2 days. I use 10% FBS with RPMI without Pen-Strep...
Hi guys, I recently experienced the same. Even 2-4 hours after seeding, cells form clumps (more than 20 cells clusters). Any solutions? Thanks!
Do know know the passage number of your cells? Have you ever harvest a cell density that is above 1 million cells/mL? I have read that a passage number over 30 and a cell density above 1 million cells does effect the cells viability in a negative way and that they won't recover.
HI EVERYONE.
After what passage number we should not use THP1 cells and why?
In my experience, one should never use THP-1 cells after passage 20-22. In fact, my transfection using the "Effectene" kit works only up to cells in the stage of passage 16. Post passage 16, the transfection efficiency gets reduced to 10 %.
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