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COS-1 cells loosely attached to culture dishes | darwyn | 2007-12-02 | Click here to register. |
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We're working with COS-1 cells (monkey kidney cells)in our lab, and lately our cells detach very easy from the culture dish surface, even by softly shaking the plates, w/o trypsin!
We buy all media already prepared from GIBCO, and also we've checked the dishes we're using are suitable for mammalian cell attachment(we use greiner bio-one 145mm culture dishes Cat. 639160)
Also, in spite of the cells appear to grow normally(except for culture medium turns from pink to yellow faster and with more intensity than it did before.
I am not sure if my cells are contaminated with either bacteria or mycoplasm, why they're then growing apparently normally?
Could anybody give me a clue. Thanks to everyone in advance.
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| trish | 2007-12-04 | Click here to register. |
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 | You might want to check your CO2 concentration in your incubator....if it even half a % too high it can cause a shift in the pH that could cause a metabolic shift that could have an effect on the ECM components that are helping the cells attach.
If the CO2 levels are "normal" then you might want to consider the contamination issue.
It has been my experience that no matter how much that I have wanted to blame the supplier, their products are well tested before they are shipped and they are rarely the cause of issues like this one. |
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| Weritab Ingo | 2007-12-06 | Click here to register. |
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| The most important part you chech on is the components of your media; ensure that it has adequate amount of L-glutamine,incubation condition are ok, while not forgetting that contaminats can affect your cells. for this use a new batch of the media and buffer solutions and see how it will go.
All the best |
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| bkeyjr | 2007-12-06 | Click here to register. |
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| I believe trish gave you the best leads. However, we use Griener dishes and we have seen similar issues with their six well plates with various lines. Check, C02 calibration, make sure you forumula is correct with your CO2 level. Then plate the same culture on several similar dishes from other vendors. Last, check for myco, but usually you will see a growth shift or doubling time shift. |
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| JAR | 2007-12-21 | Click here to register. |
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| Make sure the medium you are using rquires CO2 from the incubator. If you are using Hanks' buffered medium (.35g bicarb/L) then adding CO2 will cause the medium to quickly become yellow.
John |
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