Morphological and immunochemical studies of rat glial tumors and clonal strains propagated in culture

Abstract

✓ Rat glial tumors, induced by weekly injections of N-nitrosomethylurea for 8 months, were plated and propagated in culture. Cells grew as a glial tumor when injected back into various sites in newborn rats, and were then carried for many generations by transplantations from rat to rat or by alternate culture and animal passage. Light, phase, and electron microscopy of the cultured cells showed great variability among the glial types in all the primary tumors, in the cultures grown from them, and in the secondary in vivo tumors grown from the cultures. Many unmistakable stigmata of glia were conserved. Cloned cultures (derived from a single cell) were frozen and stored, and upon thawing resumed growth with their original histological appearance. Tumor lines from these cultured cloned strains showed much more constant growth rates and cell types; two stable lines were carried for many generations: a slow-growing astrocytoma and a faster growing glioblastoma. The distinctive neural protein called “S-100” was detected in soluble extracts of cultured cells from all five different primary tumors studied and the secondary tumors grown from them. It represented 0.2% to 0.4% of the soluble proteins extracted from the cultures, and was also present in some clonal strains and their derived tumors. One clonal strain after a few hundred generations continues to synthesize S-100. It is concluded that clonally derived cultures of gliomas histologically similar to those in man provide a stable and suitable model in rats for the study of human glia and gliomas.

Cooperation between p53 and p130(Rb2) in induction of cellular senescence

“…We also probed whether arrested cells showed any markers of differentiation, typically found in mature glial cells, 19 but could neither detect glial acidic fibrillary protein (GFAP), a characteristic marker of astrocytic differentiation, nor galactocerebroside, a marker of differentiating oligodendroglial cells. The data indicate that C6D10-p53 Val-135 adopt a senescence-like phenotype while staying in the arrest.…”

Assessment of vascular reactivity in rat brain glioma by measuring regional blood volume during graded hypoxic hypoxia

“…A cell suspension of C6 glioma (10 5 cells in 5 ml of a culture medium without foetal calf serum) was stereotaxically injected using a Hamilton syringe through a burr hole (1 mm diameter) in the right striatum (7 mm anterior to the zero ear bars, 3 mm right to the midline, 3 mm depth from the dura). The C6 glioma cell line was established by Benda et al (1971) from a methyl-nitrosourea-induced rat glioma. The C6 cells were cultured in Dulbeco's modified Eagle's medium (DMEM, GilboBRL, Lifes technologies, Scotland) supplemented with 10% foetal calf serum (GilboBRL, Lifes technologies, Scotland), 50 U ml À1 penicillin and 50 mg ml À1 streptomycin.…”

Pimonidazole binding in C6 rat brain glioma: relation with lipid droplet detection

“…Intracerebral tumours were induced by C6 cells, a cell line derived from a chemically induced malignant rat glioma (Benda et al, 1971). A measure of 5 ml of cell suspension, 10 5 cells, (prepared in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2% glutamine and 1% antibiotics) were stereotactically injected 3 mm below the dura in the right caudate nucleus of six female Wistar rats (160 -180 g) (Workman et al, 1998).…”