centrifuge at 4°C - (Dec/12/2007 )
Hello,
I just wondered why sometimes (or even always) it is recommanded to centrifuge cells at 4°C? Has anyone an idea? May be to shut down the cell metabolism?
you slow down metabolism. 4C is used to preserve the cells (and other samples), much in the same way that you preserve foods in the refrigerator.
and
when you centrifuge you generate heat (from friction, conducted from the motor, etc). refrigeration (and cold rotors) keeps the sample from experiencing elevated temperatures.
and
when you centrifuge you generate heat (from friction, conducted from the motor, etc). refrigeration (and cold rotors) keeps the sample from experiencing elevated temperatures.
Dear mdfenko,
I assume following this logic that you use PBS, Culture media and Trypsin at +4 degrees????????
Doing routine passaging of cells, they should be at room temperature.
Use +4 degrees when preparing cell pelets for enzyme preparations etc.
I was discussing centrifugation of cells today in my Institute. The ALC PK130 I use to centrifuge cells spin at 700RPM. Some groups use 1500RPM. Again in my opinion far too high as higher g/rpm damages cells.
It would be interesting to see if many other people use refridgerated (+4 oC) centrifuges as standard practice to sediment their cells
Thoughts please
Kindest regards
Rhombus
For my primary neuronal culture, we spin the cells at RT, not 4C.
When I tried to get rid of DMSO after thawing the cell line, I spun the cell at room temp.
If cell that are going to put into tissue culture flask (ie. subculture), I spin them at room temp.
If cell are for other assay (such as low cytometry, western blot), I spin them at 4 degree.
Please correct me if I am wrong.
we spin at RT, simply because the centrifuge doesn't have a temp gauge.
i actually do keep my PBS and media at 4'C. it doesn't hurt the cells, prevents bacterial contamination, and saves time. i was taught this by an RO who worked in cell culture for almost 50 years.
V
i keep my PBS , trypsin and media all in 4c but let them stand in room tepmerature for 30 minutes before starting my cell passage.
for centrifugation i do centrifuge at 4 c.
I store my media, trypsin and PBS at 4C but warm to 37C in a waterbath before using (sometimes when I'm in a hurry I'll just let them warm to about RT)
I always centrifuge my cells at RT .
I store all my media etc at 4C and then warm to room temperature or 37C, I always centrifuge at 4C and at a maximum speed of 1000rpm for 5 mins. This was how I was taught 10 years ago and as I far as I understand it, it helps preserve the cells better- like putting things on ice, any damage from DMSO or other things is slowed at the the lower temperatures. It seems from these replies that maybe it doesn't make a huge difference though.
I've done esperiments in two different labs, one has a RT centrifuge, one has a refrigerated one, cells were ok in both places!