VERO-76 Cells
VERO-76 Cells Media
DMEM pH 7.1
Calf Serum
Penicillin and Streptomycin
(Pen and Strep are optional)
500 mls
50 mls
5 mls of stock soln
Thawing VERO-76 Cells
1.
The cells are frozen in liquid N2. Be careful when handling the tubes.
2.
Thaw rapidly at 37 C in water bath.
3.
Add the 1 ml of cells to 3 mls of Vero-76 media, wait for 2 min.
4.
Add 4 mls of media and wait 2 minutes.
5.
Add 8 mls of media and plate out the cells in 1 75 ml tissue culture
flask. The DMSO will have been sufficiently diluted.
Subculturing VERO-76 Cells
1.
Once the flask has reached about 90% confluency, it needs to be
subcultured.
2.
Aspirate the media and wash the flask with 1XPBS and aspirate.
3.
Add Trypsin-EDTA at RT to the flask, about 3 mls. Rock the flask
several times until all the cells have been coated. Aspirate out most of
the trypsin-EDTA. Let flask sit for approximately 3 minutes or until
the cells start to come off.
4.
Wash the cells off with a volume of media. The trypsin-EDTA has
been sufficiently diluted.
5.
Plate out the cells at a 1:5 to 1:10 dilution
Freezing VERO-76 Cells
1.
When the cells are about 90% confluent, the cells can be frozen down.
2.
Trypsinize the cells and resuspend with 1 ml DMEM-20 (fetal calf
serum) with 10% DMSO. Freeze down 1 plate in 1 ml media.
3.
Put into cryotubes and label. Put into Mr. Frosty in -80 C freezer over
night and in the morning transfer to liquid N2.