Four permanent cell lines derived from malignant human gliomas were karyotyped using Giemsa-trypsin banding. D-65 MG had a stemline with 44 chromosomes, including 11 markers: 1p+, 2q-, 3p-, 3q+, 4p-, 9q-, 11q+, 15q-, 17p+, 21p+, 22q-. The net effect after accounting for fragments in markers was: +8, -10, -16 -X. D-32 MG had chromosome counts 90-91 without a distinct stem karyotype. Modal cells contained from 3 to 5 copies of the normal autosomes and 5 markers: 1q-, 3q-, 7q-, 13q-, 18q-. D-32 MGCl2 had a complex karyotype containing 78-82 chromosomes. There was no stemline, and modal cells varied from one another primarily in their set of marker chromosomes. A total of 23 markers were seen in this line, 17 of which were present in most modal cells. They were partially characterized as: 1q+, 1q+, 2q-, 5q+, 7p-, 7q-, 8p+, 8p+, 9p+, 12p+, 14q-, 16q+, 19q+, 19q+, a small submetacentric chromosome of undetermined origin and two small isochromosomes, i(Dp or Gp) and i(17p or 18p). A-172 MG had a modal peak of 77 chromosomes within which no two cells were exactly alike. Ten markers seen in modal cells were: 1p-, 4p+, 6p+, 6p+, 6q-, 7p-, 9p-q+, 13q+ 14p+, 22q+. There were no normal copies of chromosomes #1, #6, #9, #14. These four glioma-derived cell lines possess unique karyotypes, but each displays some combination of the numerical and structural deviations generally associated with established glioma lines.