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Message | User | Date(yyyy-mm-dd) |
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HeLa cell death, all of a sudden!! | euan01 | 2007-12-05 | Click here to register. |
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Hello All,
I have a problem and urgently need to sort it out. We have been growing Hela cells in suspension using the same procedure for nearly two years and suddenly we cannot get them to grow. This is despite regularly reviving new vials from liquid nitrogen. We make our own media and the problems seem to have arisen since we moved to this batch. Does anyone have any ideas on what we should do?
Cheers
Euan | | |
| davidj | 2007-12-05 | Click here to register. |
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 | Euan,
Can you describe how the cells are dying? Are they clumping, or just not proliferating (i.e. toxicity)?
Are you making them medium from powder? |
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| bkeyjr | 2007-12-06 | Click here to register. |
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| Make several new GLP lots, and purchase commerical forumulas. do a parrallel eval. Are all base elements the same lots as before? Dishware new or endotoxin on glassware or residual detergent? How about a new FBS lot? Do you use antibiotics? Are the antibiotics a commercial formula (perhaps you added incorrect amounts, aka toxicity)? With little info. this is all I can suggest. Testing new small lots and commerical ones against the lot in question would be the best start. FYI if you make your amino or vitamin solutions from scratch or carbs were they PH properly and in solution? Did those base elemetns have trouble filtering or the sterile filtration have any pressure? This would indicate something was not in solution and could be the issue? Just a start. |
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| JAR | 2007-12-09 | Click here to register. |
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| Check to make sure the CO2 gas is medical grade and not industrial. It could be the source of toxicity.
John |
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