I just got VERO C1008 (Kidney; African Green Monkey) and BHK-21 (C-13) (Kidney; Syrian Hamster) cell lines from ATCC. Tried VERO cell with DMEM+10%FBS+P/S first and it grew much slower compare to HEK293. It has been 5 days but only about 70-80% confluent. I wonder if nonessential amino acids and sodium pyruvate make any difference? I checked publications, some people added these two items but many people didn't. And how long it takes these two cell lines get 100% confluent? Are there any tricks about culturing these two cell lines? Would any one has hands on experience with these two cell lines provide me a detailed protocol? Any information would be very much appreciated! Thank you so much for your time!
Normally Vero cell line doesn't take more than three days to be 100% confluent (if you use 30% confluent initial cell suspension. Try DMEM High glucose, pyruvate with 3.7gm/L bicarbonate and 10% FBS, 1X P/S and 5% CO2.
BHK21 cells do fine n the DMEM (10 % FBS), though we do add non-essential amino acids and use DMEM with glutamine (glutamax). When recovering from a thaw, BHK21 cells can be a bit deceptive - we find they seem to survive at very high levels (attached to surface, normal morphology) but are quite sensitive to passaging with trypsin. I recommend giving them 3 days post thaw before trying to passage and to anticipate a significant loss of cells in that first passage.
Hi! After thawing, the cells can be slow to start back. What I do is that after thawing, I wait until I see on the microscope that a couple of cells built their extracellular matrix and are stuck to my flask (1-2 days). Since those are your healthier cells, I aspirate the media with all the thawing debris and put back fresh media without trypsinizing the couple of cells that are stuck already. This will select your healthiest cells and provide them with the proper environment to grow fast.
For VERO cells, I use AMEM media as I find they grow faster in it, but DMEM is fine too.
Hi Yanmei,
I am having some trouble with my Vero cells and hope you can give me some advice. After I thawed my cell at 37C from liquid nitrogen it grew well up but for some reasons, it stopped growing after I split it. This vero cells maintains in DMEM + 10% FBS + Pen/Strp. Also, Do you think you can spare me a few freezer stock to fest out? Thanks
After the first few passages, BHK21 cells survive passage fairly well. Don't allow them to overgrow or they will become sensitive to passage and exhibit die off. For obvious reasons, this can be frustrating when seeding experiments at specific densities.
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