DNA content and marker expression in human glioma explants

Abstract

Immunohistochemical studies of astrocytoma tissue have predominately shown fibronectin (FN) positivity restricted to vessels and glial fibrillary acidic protein (GFAP) positivity in the parenchyma. Cultured glioma cell lines, however, express both FN and GFAP. We measured the DNA content of explants of gliomas to determine if the ploidy of the FN-positive and GFAP-positive cells differed. Thirty-three explants from four high grade gliomas were cultured on slides. FN and GFAP markers were determined by double immunofluorescence. The slides were stained by the Feulgen method, the explants relocated and the DNA content measured by microdensitometry using the CAS-100 instrument. Human leukocytes applied to the slides were used as a diploid standard. Eleven GFAP-positive explants were hyperdiploid and one hypodiploid. Five FN-positive explants were diploid, three hypodiploid and ten hyperdiploid. One FN-positive explant was biclonal with aneuploid subpopulations. Two hyperdiploid explants, each of which had monoclonal histogram patterns, expressed both FN and GFAP. We conclude that most FN-positive cells, in addition to GFAP-positive cells, from cultured gliomas represent neoplastic cells. These may be present in the tumor in low numbers or may result from marker switching in culture.