primary cells better than 3T3 cells? - (Dec/16/2006 )
Hello,
I need to prove my in vivo gene expression results and as all the protein detection methods are available are not working for my proteins (e.g. protein concentration to low in tissue extract). Therefore my supervisor told me to use 3T3 cells, as we would like to study the expression in fibroblasts. I am not very successful growing primary skin fibroblasts or synovial fibroblasts so far and I need results, thats wy I want to use a cell line.
But do 3T3 represent my in vivo system? Somebody told me, they produce everything, thats why they are no good?
Or should I order primary cells?
I need to prove my in vivo gene expression results and as all the protein detection methods are available are not working for my proteins (e.g. protein concentration to low in tissue extract). Therefore my supervisor told me to use 3T3 cells, as we would like to study the expression in fibroblasts. I am not very successful growing primary skin fibroblasts or synovial fibroblasts so far and I need results, thats wy I want to use a cell line.
But do 3T3 represent my in vivo system? Somebody told me, they produce everything, thats why they are no good?
Or should I order primary cells?
not easy to answere; if you analyze distinct parts of signal transduction pathways in 3T3 they may represent native fibroblasts; however, immortal cell lines differ in many aspects, especially in biological function and inner regulation; 3T3 overexpress or suppress expression of some genes in comparison to fibroblasts; so, if you need a set of certain proteins in the cell for your study, you may not hit your biological question if you use 3T3;
try to find published references which may help to decide to use 3T3 or native fibroblasts
if you want human cells, IMR90 cells are suitable for analysing proteins.
Nuclear extracts of these cells are not very good for in vitro transcription assays, as observed in my lab.
IMR90 are human primary fibroblasts. We get ours from ATCC
3T3-L1 cells produce triglycerides and undergo pre-adipose to adipose like conversion as they progress to a contact inhibited state. They are good transfection host using fugene.
If you think this cell type is not important for your work, you might want to use normal human fibroblasts like MRC-5 or WI-38 which can be grown in culture for up to 30 passages.
Hope this helps.