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Message | User | Date(yyyy-mm-dd) |
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Problems with ATCC cells | joanksza | 2001-11-20 | Click here to register. |
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Has anybody had problems with viability of cells delivered from ATCC? We thawed 4 vials out of 10 and there wasn't a single viable cell. Unfortunately warranty period passed and probably we will have problems with receiving new cells.
If anybody had the same problems please contact us and write what did you do. | | |
| jpartee | 2001-11-26 | Click here to register. |
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 | Cells from ATCC are shipped with a high number of viable cells. The reason you are getting problems can be from several factors.
1. The cells lost viability due to incomplete cooling in the dry ice that they were shipped in. Cells left to long in dry ice lose viability rather quickly.
2. If you get the cells and store them at -80c and not in liquid nitrogen, they lose viability over time. You can expect a loss of about 10% or more per day in a -80C freezer. Some cells last longer but don't count on it.
3. The media you used was different than the recommended media that they where originally grown in. If you change media on a cell without proper adaptation time, they will go into "shock" and either die or not grow for long periods of time. If it is a hybridoma, they look dead when thawed and placed in media. You need to keep them for at least 48 hours in media before they start to grow. You cannot tell viability with just a microscope, usually.
The most common problem is a bad FBS lot or antibiotic toxicity. The best cells to get are serum-free cells since you can eliminate the FBS variable.
It is better to NOT use antibiotics if you can help it. Some cells don't like them. If you are sure of your clean room and sterile procedures, nix the antibiotics.
It is possible that you got a bad lot of cells but the quality control at ATCC is very good, they don't want to have to keep replacing dead cells (costs lots of money) so they do everything they can to get them to you ready to go.
Remember, ATCC has been around for over 70 years.
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