In situ cell kinetics studies on human neuroectodermal tumors with bromodeoxyuridine labeling

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✓ Thirty-eight patients undergoing surgical removal of neuroectodermal tumors of the central nervous system were given a 1-hour intravenous infusion of bromodeoxyuridine (BUdR), 150 to 200 mg/sq m, to label tumor cells in the deoxyribonucleic acid (DNA) synthesis phase (S-phase). The excised tumor specimens were divided into two portions: one was fixed with 70% ethanol and embedded in paraffin and the other was digested with an enzyme cocktail to make a single-cell suspension. The paraffin-embedded tissues were stained by an indirect peroxidase method using anti-BUdR monoclonal antibody (MA) as the first antibody. Single-cell suspensions were reacted with fluorescein isothiocyanate (FITC)-conjugated anti-BUdR MA's for flow cytometric analysis. S-phase cells that had incorporated BUdR into their DNA were well stained by both methods. The percentage of BUdR-labeled cells, or S-phase fraction, was calculated in tissue sections by microscopic examination and in single-cell suspensions by flow cytometric analysis. The biological malignancy of the tumors was reflected in the S-phase fractions, which were 5% to 20% for glioblastoma multiforme, medulloblastoma, and highly anaplastic astrocytoma, but less than 1% in most moderately anaplastic astrocytomas, ependymomas, and mixed gliomas. Two juvenile pilocytic astrocytomas and two low-grade astrocytomas from children had high S-phase fractions despite the fairly benign and slow-growing nature of these tumors. These results indicate that the S-phase fraction obtained immunocytochemically with anti-BUdR MA's may provide useful information in estimating the biological malignancy of human central nervous system tumors in situ.

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Address reprint requests to: Takao Hoshino, M.D., c/o The Editorial Office, Department of Neurological Surgery, 1360 Ninth Avenue, Suite 210, San Francisco, California 94122.

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Figures

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    Photomicrographs of a juvenile pilocytic astrocytoma of the cerebellum in Case 30. × 200. Left: Alcohol-fixed tissue section. H & E. Right: Section prepared with immunoperoxidase stain for bromodeoxy-uridine (BUdR), counterstained with eosin. Many proliferated endothelial nuclei are BUdR-positive (nuclei stained dark).

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    Photomicrographs of a pineocytoma in Case 38. × 600. Left: Homer-Wright rosettes and a perivascular rosette are visible. H & E. Right: Many bromodeoxyuridine-positive tumor nuclei are shown on a section without counterstain.

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    Photomicrographs of a highly anaplastic astrocytoma in Case 8. × 190. Left: Alcohol-fixed tissue showing high cellularity and pleomorphic nuclei. H & E. Right: Immunoperoxidase stain for bromodeoxy-uridine, without counterstain, shows many labeled tumor nuclei.

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    Contrast-enhanced computerized tomography scans of Case 8, taken before (left), 1 week after (center), and 50 days after (right) initial tumor removal.

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