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Difficult Cell Proliferation Experiment


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#1 Baars01

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Posted 05 April 2011 - 01:30 AM

Hi,

I would like to assess cell proliferation of a cell type after siRNA mediated knock-down of certain genes. Depending on the knock-down these cells can differentiate and senesce. I would like to measure the cell proliferation of these cells after transfection, but the differentiation program only comes into full swing after three days. This is unfortunately also the time at which the cells are confluent, so any proliferation assay beyond three days is difficult to do as the cells have to be harvested, counted and reseeded to a new plate. This introduces variance in cell numbers between the wells which skews any MTT or WST-1 data.

Would anybody have any ideas on how to set up this experiment. I have thought of live cell observation and imaging over time, but any simpler methods would be very welcome

Thanks

#2 bob1

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Posted 05 April 2011 - 03:30 PM

Have you tried seeding and transfecting the cells at lower density?

Are the cells contact inhibited?

#3 Baars01

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Posted 06 April 2011 - 12:15 AM

Have you tried seeding and transfecting the cells at lower density?

Are the cells contact inhibited?


Unfortunately the cells are already at the lowest density possible for their type




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