We were successful in transforming 3T3 cells by murine herpes virus MHV-68. The cells acquired the typical "criss-cross" pattern of growth" character and were morphologically different from original 3T3 cells. However, several cells detached, remained viable and formed colonies even in a liquid culture fluid. Have someone experience with such cell transformation?
Sometimes, transformed cells can lose their adhesion-dependent status and can grow as floating cells. The phenomena may be a consequence of unexpected gene mutation or deletion.
Besides, 'colony formation in floating status' does not really mean 'colony formation from monoclonal descendants from single cell.' Like lymphoid cell lines, floating cells can form clumps. So if need to clone. such 'floating transformant' you have to repeat limited-dilution method.
Hi, Franz
I also use MHV68 but in my hand this virus usually killed the infected cells. I didn't know that MHV68 can transform cells.
Anyway, transformed cells may show anchorage independent growth. I didn't have the experience but I guess you may want to try soft agar assay and nude mice xenograft model.
Hi, Jacky
Thank you for your comment. I remember that rodent derived primary cells including NIH3T3 cells are transformed quite easily. Such spontaneous transformation occurs frequently even without any infection nor transfection. So, I recommend Franz should forget about the colonies and transformants unless otherwise he needs to analyze the latent status of the virus.
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