do not need to do this, but you should leave the spleen at maximum 10 minutes after killing the mouse and Remove the spleen cells at a maximum of 3 times by slow infusion with PBS or serum free culture media.
Thanks Reza Hadavi for the suggestion. Could you please elaborate the procedure for culturing the spleen cells after removal from the mouse? Or can you cite any article for that?
The production of hybridoma requires a lot of care and repetition., as there are plenty of key points in this work. So before doing the original fusion, first do a test fusion with an unimmunized mouse.