I am having problem with monoclonal antibody. The fusion partner I am using is SP2/0-ag14 (ATCC:1581) and I am growing it in DMEM+10%FBS+ Penicillin:streptomycin. The SP2/0 seems normal (round suspension cells) until I perform fusion! After I performed fusion and added HAT medium, the morphology of the cells change to fibroblast-like (please see attached pictures!) I am sure this strange cells are Sp2/0 as I have encounter this before in another lab and fusions using NS0 fusion cell lines do not produce this strange cells! Anyone know what's wrong with my SP2/0????? This strange Sp2/0 is surrounding my hybridoma cells in EVERY wells and I am suspecting it may be the reason why I have only little hybridoma cells growing! (none of them is positive!!!!!)
Please help me as I am really struggling to get a monoclonal antibody! (I cant change the fusion partner cell line as I dont have money to buy a new one ><) Is this strange morphology normal to Sp2/0???? (I have seen it before in another lab) or is it because something wrong with my medium? but my sp2/0 grow well in dmem usually, just except after fusion that it go strange! I suspect the HAT medium as well but normal unfused SP2/0 just die so I dont know where this strange cells come from?
Any advise are greatly appreciated! Thanks a lot in advance!
Edited by vivianchan, 29 August 2010 - 11:31 PM.