monoclonal cell line vs. polyclonal cell line

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Death Adder
Death Adder
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Joined: Fri Sep 23, 2005 7:58 pm

monoclonal cell line vs. polyclonal cell line

Post by kk » Wed Mar 01, 2006 1:52 am

I am so confused. If I transfected cells, and selected for stably transfected cells by adding selection antibody for ~12 days (or till the untransfected=control cells die), than I have a polyclonal stably transfected cell population. Right?

If I take this "polyclonal cell line" and isolate individual cells (by serial dilution or by a sophisticated machine) and subculture them, than I will get "monoclonal cell line". Right?

Question 1:

for further assays (real time, Western, migration assay) is it really crutial to use the "monoclonal = one-cell-derived cell line" or it's fine to use the "polyclonal"? WHY?

My argument: if I have 3 "monoclonal cell lines" than I have to do all the assays (in triplicates) using all three clones? It's already 9x the 1 assay and a lot of time for the clonal seletion.

Question 2:

using a "monoclonal cell line" really does show the effect of the actual trangene instead of the position-effect of the integration?

My argument: I don't think so, even if I work with 3 different clones. I think the overall effect of the transgene is MORE represented by the "polyclonal cell line" where all the cells bear the extra information but the integration site-position-effect is more masked.

Please, write your opinion.

P.S. I see numerous publications in high impact journals where they simply used the "polyclonal cell line" right after the antibiotics-selection...and I think it's fine.

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