I am trying to differentiate PC12 cells in 96 well plate ( 10000/ well) with the help of NGF but my cells kept on dividing even after the addition of NGFand forms a clump so they do not differentiate properly. Can anyone suggest a protocol with NGF?
Nitish,
You might have to serum starve the cells when you add NGF and do not change medium until it is differentiated completely. If you realize cells need more medium, just add more to the existing medium. Get down the serum to 2 % and add NGF containing medium while passage and adding cells into the 96 wells. Also do not shake the plate after adding cells for at least 20-30 min until all cell stick to bottom.
Good luck !
Hi Nitish,
I have been trying to use NGF for differentiating PC12 cells and am having difficulties in differentating them. They are still dividing as you said. I have also tried using different NGF concentrations (50-200 ng/ml) and the cells still continue to divide. It seems that the cells begin to differentiate but I am not able to see long processes. Do you have any suggestions?
Thank you.
Khushboo.
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