Re: Sorting Hybridoma Cells

Mike Clark (mrc7@mole.bio.cam.ac.uk)
Wed, 26 Jul 1995 12:36:57 +0059 (BST)

During my PhD project in Cesar Milsteins laboratory in 1978/1981 I
conducted experiments in which I fused myeloma cell lines with B-lymphoma
cell lines. Obviously the B-lymphoma cell lines had large amounts of
detectable surface immunoglobulin but very little secreted immunglobulin
was detected. After cell fusion the hybrid cells secreted the
immunolgobulin in large amounts and at the same time there was a drop in
surface immunoglobulin.
During prolonged culture some of the hybrid cell lines gradually lost the
ability to secrete the immunoglobulin and the level of surface
immunglobulin rose again. So in this example sorting for the highest
levels of surface immunoglobulin would not give the highest levels of
secretion.

Now the question arises as to how these results compare to fusions with
spleen cells? It is my opinion that the myeloma cell is able to fuse to
and immortalise B-cells at various stages of differentiation and that the
myeloma cell can also induce the secretion (through changes in mRNA
processing) of immunoglobulin which may have been predominantly cell
surface on the parental B-cell. Alternatively if the parental cell were a
plasmacytoma there may have been very little surface immunolgobulin to
start with. If you are interested in sorting for immunoglobulin secretion
probably the best assay is for secretion! The encapsulation techniques
suggested by others on this mailing list sound ideal.
Regards,
Mike Clark, mrc7@cam.ac.uk http://www.path.cam.ac.uk/~mrc7/

--
  o/ \\    //            ||  ,_ o   Dr. M.R. Clark, Division of Immunology
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