Protocol Online logo
Top : New Forum Archives (2009-): : General Lab Techniques

Difficult Cell Proliferation Experiment - (Apr/05/2011 )

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

-Baars01-

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

Are the cells contact inhibited?

-bob1-

bob1 on Tue Apr 5 23:30:57 2011 said:


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

-Baars01-