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Inconsistent results with retroviral transduction

retroviral transduction t cells gfp retroviral vector

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#1 NovaCroft

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Posted 19 January 2015 - 06:18 PM

Hi all,

 

I ve been doing retroviral transductions on t cells for awhile now and while i get transduction efficiencies of 50% or more, the results i get from the experiments i set up with the transduced cells vary from experiment to experiment.

To be more specific, i analyze the effects of over expressing a protein on t cell proliferation. most of the time it reduces proliferation but of late it doesnt do anything to proliferation. I know its not because of a mutation in the vector or lack of protein expression. In my western blots, its obvious the protein is clearly over expressed.

Any thoughts on what could be going on here would be appreciated!

 

Cheers!



#2 CPRES

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Posted 20 January 2015 - 12:56 PM

So many things can be going on..

 

1. You have (unwittingly) changed methods, reagents, cells, virus, etc.

2. You are not getting the same (high or low) protein expression levels.

3. Previously, your experiments were killing off your cells, now you have improved.

4. Your proliferation assay has changed. for example: you were/are counting cells wrong.

 

I would suspect and troubleshoot technical parts first, before agonizing over possible scientific explanations, which I have none to offer.


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Also tagged with one or more of these keywords: retroviral transduction, t cells, gfp retroviral vector

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