Generating a low-maintenance transformed cell line

Tony Schountz tschount at mesastate.edu
Wed Feb 28 17:42:30 EST 2001


I'd like to generate lymphoid cell lines from an unusual, outbred mouse
species. There are no known transforming viruses for this species and
I'd like to avoid carcinogens. What I think would be best would be to
clone some cell cycle gene (or genes) into a mammalian expression vector
(e.g. pCR, pTriEx, etc.) and then stabley transfect spleen cells.
Ideally, no growth factors would be required; routine FBS-containing
media (e.g. RPMI, DMEM) would be all that the cells need. If possible, a
lab mouse (Mus musculus) gene would be ideal (as opposed to some viral
oncogene).

I have tried generating xenoreactive T cells from these mice using
BALB/c (and a couple of other lab mice) stimulators, but they do not
seem to recognize the MHC molecules well enough to survive as lines,
even though the T cells are responsive to human recombinant IL-2. I
haven't tried generating T cell lines to minor antigens.

The use of these cells would include long-term sources of (1) genomic
DNA, RNA and proteins, (2) monocytic, B and T cell lines for rudimentary
leukocyte assessment (not necessarily Ag-specific), and (3) for
screening hybridomas for monoclonals reactive to cell surface antigens.

Any suggestions as to what gene(s) would be good candidates for this
proposal? Or perhaps alternative strategies?

Thanks,

Tony

--
Tony Schountz, Ph.D.
Department of Biological Sciences
Mesa State College
mailto:tschount at mesastate.edu







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