Concerning numeration cells passage, there are two school of thought. One said that we have to restart from zero and one said that we have to continuing the numeration (e.g. freeze at #4, maintain at #4). Could anyone provide me a precise reference? I searched on the web but I didn't find.
Thank you in advance!!!
Eleonora
Of course you should continue incrementing the passage number! Freezing the cells does not extend telomeres, and does not reprogram the cells to their original state, before selection / culture stress; on the contrary it contributes to all this stress. I'm afraid I don't have any reference for this, but it is rather obvious I think, unless I misunderstood your question.
Hi,
This I have got from a reference, which is as follows,
"Cells are frozen at the lowest generation number possible and used to replace stocks at regular intervals before the onset of senescence. When thawed the generation number resumes at one over the number at freezing. For continuous cell lines, the number of passages since last thawed from the freezer is recorded. If the passage level at freezing is known, this may be added on but often this is not known."
the reference is-http://www.nature.com/bjc/journal/v111/n6/full/bjc2014166a.html
I think the passage number is calculated from the first subculture of the cell line and should be kept in mind after freeze-thawing cycle. That is why it is recommended to keep many frozen vials of cells at low passage number, so that you can use them whenever the cell passage number of the current culture becomes too high and you can discard the old cells.
Therefore, as you said a vial frozen at 4th passage will be thawed at the 4th passage and the next subculture would be 5th passage and so on.
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