I want to do some experiments on Kupffer cells. I have done it in vivo. However, it's a little difficult to isolate and culture Kupffer cells for me. My question is can I use RAW264.7 to repalce Kupffer cells for my in vitro study?
What will the reviewers challenge if I use RAW264.7 cells?
Thanks for your attention.
RAW264.7 is a mouse NF-kB secreted alkaline phosphatase SEAP - (SEAPorter) stable reporter cell line, which expresses all kinds of Toll-like receptor other than TLR5. This cell line is widely used as mouse macrophage in vitro, but not as specific to liver. Macrophages distribute in the several organs, as you know well, in the brain, skin, liver, lungs etc, and each organ affects the phenotype of the differentiated and specialized macrophages via the cell-to-cell interaction, cytokines, chemokines and hormones. That is why I do not think RAW264.7 is not the appropriate material to perform experiments for Kupffer cells. Instead, I strongly recommend to use immortalized rat Kupffer cells with SV40 antigen, which you can purchase from COSMO BIO (http://www.cosmobio.co.jp/connections/p_ku_view.asp?PrimaryKeyValue=9487593&ServerKey=Primary&selPrice=1) or isolate from the liver and perform ex vivo experiments.
http://www.cosmobio.co.jp/connections/p_ku_view.asp?PrimaryKeyValue=9487593&ServerKey=Primary&selPrice=1
Hello
You are right about their limited popularity. This is mainly due to previous unavailability of good quality primary Kupffer cells or difficulty with their preparation as in case of fibroblast culture (need for micro-patterning or careful seeding techniques). At least in the first case, Life Technologies currently offers good quality preps of cryopreserved rat Kupffer cells (http://products.invitrogen.com/ivgn/product/RTKCCS?ICID=search-product) that can be readily plated together with primary hepatocytes to build inflammatory co-culture. Those cells has been extensively tested for purity, function and show the same cytokine response after LPS stimulation as corresponding freshly isolated Kupffer cells. In our initial tastings, those cells perform significantly better than current macrophage cell lines (for example SV40 , RKC1, RKC2 and RAW264.7) you have good choice, but i agree with Go J Yoshida - , the best of these cell lines for in vitro study is SV40
Good luck
Hey,
Another possibility, though not specific for Kupffer cells, is that you test your effect on other macrophage-like cells. F.e. bone-marrow derived macrophages or peritoneal macrophages (both are relatively easy to isolate). I don't know exactly what you're investigating, but this can give you information what the effect are on macrophages
Good luck
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