Open AccessCell kinetic studies of in situ human brain tumors with bromodeoxyuridine
Author(s) -
Hoshino Takao,
Nagashima Tadashi,
Murovic Judith,
Levin Ellen M.,
Levin Victor A.,
Rupp Stephen M.
Publication year - 1985
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990060619
Subject(s) - bromodeoxyuridine , flow cytometry , microbiology and biotechnology , staining , monoclonal antibody , chemistry , cell cycle , dna , horseradish peroxidase , in situ , cell , pathology , biology , antibody , enzyme , biochemistry , cell growth , immunology , medicine , organic chemistry
Abstract At the time of surgery, 18 patients with various brain tumors were given a 1‐h i.v. infusion of bromodeoxyuridine (BrdUrd), 170–200 mg/m2. At an infusion rate of 200 Mg/M2/h, serum BrdUrd levels of 8 μM were achieved. After the infusion, tumor tissue was obtained and divided into two portions. One portion was fixed in 70% ethanol, embedded in paraffin, and sectioned; the sections were deparaffinized, denatured with 2 N HCl, and reacted with monoclonal antibodies against BrdUrd (anti‐BrdUrd MAb). BrdUrd‐labeled nuclei were demonstrated satisfactorily by an indirect peroxidase method. The other portion was dissociated into single cells with a DNase enzyme cocktail and reacted with FITC‐conjugated anti‐BrdUrd MAb to determine the percentage of BrdUrd‐labeled cells or with chromomycin A3 for DNA analysis. The single‐cell suspensions were analyzed by flow cytometry. The fraction of S‐phase cells in the tissue sections was similar to both the percentage of BrdUrd‐labeled nuclei and the S‐phase fraction determined by flow cytometric analysis. The results obtained with BrdUrd‐labeled nuclei were similar to those obtained from previous autoradiographic studies of various brain tumors exposed to a pulse of 3 H‐thymidine. Since BrdUUrd is not radioactive and is nontoxic at the dosage used, these techniques, together with the histopathological diagnosis, may help to predict the biological malignancy of individual tumors.