inhibitory effect of macrophages - (Mar/11/2007 )
i am facing a problem regarding monoclonal antibody production. we used macrophages( feeder layer) from a mouse which had been already immunized with some unknown immunogen 6-8 months back which might have led to the formation of ascites, however we continued using it for fusion . we observed the death of cells on the 3rd day of fusion and formation of clumps. can anyone suggest a reason for this !!!!!!!!!!!!!! we assme it to be due to the inhibitory effect of macrophages.
if the mouse has been used for some other immunisation or not should be of no consequence, since the macrophages (peritoneal, i assume) are part of the innate immune system and have therefor nor immunological "memory"... that is to say, macrophages from immunised mice and from naive mice should not be different. and when you use feeder macrophages, normally everything should be ok... but maybe you want to consider some infection of your mice that you transfer to your fusion cells via the macrophages?
i dont want any infection of the mice .. i just wanted to know why hybridomas seen after some days of fusion started to die???? morphological wise it looked like hybridomas.. it grew in HAT????
i want know one more thing.. whether ascites have any inhibitory effect ??????????