Part VI: Freezing and viability staining of cells
Welcome to the last of a series of articles providing useful hints for culturing
animal cells. This article contains considerations for freezing and viability
staining of cells.
For some cell cultures, especially valuable ones, it is common practice
to maintain a two-tiered frozen cell bank: a master and a working cell bank.
The working cell bank comprises cells from one of the master bank samples,
which have been grown for several passages before storage. If and when future
cell samples are needed, they are taken from the working cell bank. The
master cell bank is used only when absolutely necessary. This ensures that
a stock of cells with a low passage number is maintained, and avoids genetic
variation within the culture.
1. Check that cells are healthy, not contaminated, and have the correct
2. Change the medium 24 h before freezing the cells.
Tip - Adherent and suspension cell cultures should not be at a high density
for freezing. We recommend freezing cells when they are in the logarithmic
3. Adherent cultures: harvest the cells by trypsinization, resuspend in
medium containing serum, centrifuge at 200 x g for 5 min, and then resuspend
cells in freezing medium (see Table 1) at a density of 35 x 106 cells/ml.
Suspension cultures: centrifuge the cells at 200 x g for 5 min, and resuspend
in freezing medium (see Table 1) at a density of 510 x 106 cells/ml.
Freezing medium containing DMSO is hazardous and should
be handled with caution.
Source:Page: All 1 2 3 Related biology technology :1
. The QIAGEN Guide to Animal Cell Culture2
. The QIAGEN Guide to Animal Cell Culture3
. The QIAGEN Guide to Animal Cell Culture4
. The QIAGEN Guide to Animal Cell Culture5
. The QIAGEN Guide to Animal Cell Culture6
. QIAGEN Instrument Service insist on the best in service and support7
. QIAGEN Multiplex PCR Handbook8
. QIAGEN Multiplex PCR Kit9
. QIAGEN Plasmid Kits10
. Laser microdissection and nucleic acid purification - a Leica - QIAGEN
. QIAGEN PCR CloningPlus Kit