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Message | User | Date(yyyy-mm-dd) |
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Use of trypsin inhibitor in primary cell culturing | An | 2009-03-18 | Click here to register. |
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Hi!
For my research I am coculturing neurons on top of astrocyte monolayers in serum free medium. However, the plating of primary neurons on top of the astrocytes does not lead to consequently good results. Sometimes the astrocytes start to deform (they become stringy en start to form "holes") and ultimately the cells will die!
For the culturing of the neurons, I dissociate hippocampal tissue in a trypsin solution and wash this solution with serum free medium. In our protocols we never use trypsin inhibitors, but I have now some very strong indications that the trypsin might be damaging my cells :s
So, I am considering to use trypsin inhibitor. But I would like to know if there might be any cons to do so? (Otherwise I just have to start washing the tissue better and more).
Any advice/help/information on this subject will be greatly appreciated!!!
Thx!! | | |
| Grifta | 2009-03-23 | Click here to register. |
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 | Try washing the tissue in media with serum in, as this inactivates trypsin.
Paul |
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| RTE | 2009-03-25 | Click here to register. |
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| Also the lack of serum could be causing problems with the attachment of the primary neurons. A small amount of serum would help this or a specialty serum free medium that contains attachment factors might help. |
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