THP-1 cell culture - (Dec/12/2008 )
Hello everyone!
I'm planning to work with THP-1 cells but I don't know anything about cell culture. I would like to know if anyone can tell me where can I find a detailed protocol of how to culture these cells.
Here you will find recommendations from ATCC
THP-1 on ATCC webpage
start with 0.3x10^6 cells per mL and don't let them grow over 1.10^6/mL
good luck
I'm a bit confused because I'm new in the cell culture, so I will ask a question that might sound a bit strange. I've been reading the forum on how to revive THP-1 cells. Do I have to follow this protocol when I receive the cells directly from the company where I buy them? What I mean is, do cells come in cryovials or not? I've read the ATCC's and the Sigma's information about this cell line but I'm still not clarified on the culture procedures.
Another thing is the RPMI medium. ATCC recommends their own RPMI which has already L-glutamine included but I read that L-glutamine is unstable so it should be added to the medium instead of being included in it. Does anyone recommend another RPMI? Has someone used RPMI from Sigma?
Another thing is the RPMI medium. ATCC recommends their own RPMI which has already L-glutamine included but I read that L-glutamine is unstable so it should be added to the medium instead of being included in it. Does anyone recommend another RPMI? Has someone used RPMI from Sigma?
I would recommend you RPMI plus glutamax ( eg invitrogen). It's an other form of glutamine, but more stable.
I would say they will come in cryovial ....
thaw the cells quickly ( in a clean water-bath at 37 degree celsius, but I prefer in the hand, because water-bath are not clean) while there is only a little ice in the tube, clean it with ethanol 70 % before to open under hood, dilute the cells in pre-warmed medium, centrifuge and resuspend in fresh medium at the right concentration. I realize their is some discrepancy between my recommendations and ATCC ones. It seems you can dilute more that what I said. However, cells do not like to be too much diluted. If cells don't start to grow, you should concentrate them a little more.
When you dilute the cells, you should count them with trypan blue.
You should try to find someone in the institute who could help you with cell culture because there are various things that one should show you and we can't explain you by mail.