1). Wound healing
2). Cell proliferation
3). Apoptosis
4). Etc...
I have A549, MCF7, HepG2, THP1 .
So please suggest to me, which passage number is suitable for above assays?
My work : pcDNA3.1+Insert+GFP (Expression studies)
in my opinion yes- you basically can't avoid clonal selection due to extensive passaging the cells. For cancer cell lines, I would not go beyond passage 30.
I would stay in the same range if you like to keep your experiments compareable.
as a general rule of tumb I always go for as low as possible (to have enought frozen stocks acquired) in passage number.
good luck!
in my opinion yes- you basically can't avoid clonal selection due to extensive passaging the cells. For cancer cell lines, I would not go beyond passage 30.
I would stay in the same range if you like to keep your experiments compareable.
as a general rule of tumb I always go for as low as possible (to have enought frozen stocks acquired) in passage number.
good luck!
I would be very cautious about comparing high and low passage numbers for wound healing and proliferation assays.
With each passage, you run the risk of unintentionally selecting for more adherent cells, especially if you re-use the same flask between passages. This could affect the wound healing assay because increased adherence could affect migration/metastasis (Leung-Tack et al Int J Cancer. 1988).
With each passage, there is also the risk for clonal expansion of cells that proliferate faster (Hughes et al, Biotechniques. 2007), which could eventually take over your cell line. Therefore cell proliferation assays should be done on cells with similar passage numbers.
Thank you very much for your valuable suggestion...
I have experience in studying genetic aberrations of leukemic and breast cancer cell lines before. In the study, we have found that cancer cell line with passage number more than 15 have got chromosome and gene changes. This really an interesting question. I wonder if there any articles which may support your point Elisabeth Stein and Fen Li besides information that have been given by Jason Higa.
There are articles, I will only cite 2 of them.
This one deals with CHO cells:
http://www.ncbi.nlm.nih.gov/pubmed/22331235
and a review dealing with this issue:
http://www.ncbi.nlm.nih.gov/pubmed/18072586
best regards
Elisabeth
According to me, Yes! You will have to perform all assays within more or less similar PN.
Elisabeth Stein , i doesn't have Springerlink access please send this article" Effects of high passage cultivation on CHO cells: a global analysis"
Thank You Elisabeth Stein... i will use beyond 30 passage.
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