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Fig. 5. Pre-neoplastic cells can be isolated from the cerebellum of
patched mutant mice. Cells were isolated from the cerebellum of
wild-type and patched+/– mice by enzymatic
dissociation followed by Percoll gradient centrifugation, and viable cells
were counted after Trypan Blue staining. (A) The average yield for 6-week-old
wild-type mice was 0.53±0.25 million cells. For patched
heterozygotes of the same age, the average yield was 4.3 million cells, with
84% of animals having more than 0.9 million cells (the maximum number seen in
wild-type mice). (B) Among older patched mutants (10-25 weeks), 16%
had tumors containing 50-600 million cells; the remainder had fewer than 2
million cells. (C) Non-granule cell precursors (GFP- cells from
neonatal Math1-GFP/patched+/– mice), pre-neoplastic
cells and tumor cells were stained with the fluorescent ß-galactosidase
substrate FDG and analyzed by flow cytometry. Relative fluorescence of
non-GCPs (blue), pre-neoplastic cells (pink) and tumor cells (purple) is
shown. The horizontal black line indicates the range of fluorescence
considered to be positive (i.e. above background); 89% of pre-neoplastic cells
and 96% of tumor cells exhibited fluorescence within this range.
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