Hi!
I would like to know how cell passage number affect cells. Early passages are good for freezing.
What would happen if cells are kept passaging continuously ?
Thank you.
Dear Sujani,
at high passage numbers, cells experience changes in morphology, growth rates, protein expression and transfection efficiency, compared to lower passage cells. The use of cell lines that have been in culture too long should therefore be avoided to ensure reliable and reproducible results. There is no general method for determining the optimum passage number of a cell line because effects are complex and vary with the cell line, tissue and species of origin, culture conditions and the application for which the cells are used.
May I recommend that you monitor over time (in a control chart) certain characteristics of your cell line significant for your application. In dioxin analysis, we use parameters of our TCDD-standard curve (we expose genetically modified hepatoma cells to TCDD and measure their response) such as the EC50 value and the fold induction at a certain (low) TCDD-concentration. When those fall above or below certain values, we discard the cell line and start culturing new cells from our stock. Another option, if you do not do such or other measurements, may be certain changes in morphology visible under the microscope. Just take the courage to establish your own criteria and limit values, based on your experience. And if you are a beginner, it is even more fun as experience will grow.
More interesting information is found here:
https://www.atcc.org/~/media/PDFs/Technical%20Bulletins/tb07.ashx
Hope this helps. Wishing you best of success,
Johannes
Hi Johannes, thank you so much for your answer with clear explanation.
I second Dr. Haendrich. For instance, some adherent cell lines are expected to be used at high passage levels to take advantage of certain morphological or physiological characteristics. I've worked with Caco-2 cell line, at high passage values (>30), because this cell line (human colorectal cancer) it is expected to differentiate into epithelial cells when reaching several passages. In addition, you might also verify the genetic and microbiological integrity of your cell cultures using suitable tests to guarantee if they still have those characteristics from their own cell line.
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