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hybridomas

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hybridomas

Postby xbritoe » Mon Feb 06, 2012 5:26 am

Hi, I was wondering does anyone have experience working with hybridomas? This is my first time working with them and I notice that in my 6-well plate I see a lot of little white dots, and when I look under a microscope I see circular shaped cells. I just wanted to make sure that those are the hybridomas and that its normal to be able to see the white dots in the culture by just looking at it with your eyes. I'm use to working with adherent cells and suspensions cells so typically the cells aren't as visible unless you use a microscope. I just want to make sure that there is no contamination within my culture. Thanks.
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Postby biohazard » Mon Feb 06, 2012 7:23 am

I cannot say for sure without seeing your white dots, but many suspension cells grow in "clumps", meaning that a cell has started to divide there and has generated a lot of daughter cells at that location. Another possibility is that the cells simply stick into one another, causing visible clumping.

Do the "white dots" disappear if you gently mix them with a pipette? If so, they are most likely just clusters of hybridoma cells - if there are enough cells in the cluster, you can see them by naked eye.

Other possibilities are precipitates from your culture medium (you should then see crystals of some kind and they would not disappear easily by mixing) or some other contamination (e.g. bacteria, this should be obvious in most cases since the culture medium becomes turbid, the pH changes quickly and the cells soon die).
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