Cell kinetics of rat 9L brain tumors determined by double labeling with iodo- and bromodeoxyuridine

A. Asai, S. Shibui, M. Barker, M. Vanderlaan, J. W. Gray, T. Hoshino

Research output: Contribution to journalArticlepeer-review

28 Scopus citations


Rats with 9L brain tumors received intraperitoneal injections of iododeoxyuridine (IUdR) and bromodeoxyuridine (BUdR) to estimate the duration of the deoxyribonucleic acid (DNA) synthesis phase (Ts) and the potential doubling time (Tp) of individual tumors. Different sequences and intervals (2 or 3 hours) of IUdR and BUdR administration were evaluated. After denaturation, tumor sections were reacted with Br-3, a monoclonal antibody that identifies only BUdR, and then were stained immunohistochemically by the avidin-biotin complex method. An antibody that recognizes the IUdR and BUdR, IU-4, was applied to the sections and identified by the alkaline phosphatase-antialkaline phosphatase method. Nuclei labeled only with IUdR stained blue, while those labeled with BUdR or with BUdR and IUdR stained brown. The fraction of cells that either left or entered the S-phase during the time between administration of IUdR and BUdR was measured to calculate Ts and Tp, assuming that the labeled cohort completed the DNA synthesis at a constant rate. The Ts was 8.8 hours (coefficient of variation (cv) = 0.05) and the Tp was 64.2 hours (cv = 0.08). The sequence and interval of administration of IUdR and BUdR had a minimal effect on Ts and Tp. In studies of 9L cells in monolayer culture, the Ts was 9.6 hours (cv = 0.08) and the Tp was 30.6 hours (cv = 0.06). Double labeling with BUdR and IUdR allows the duration of the S-phase and potential doubling time of individual brain tumors to be estimated in situ from a single biopsy specimen.

Original languageEnglish (US)
Pages (from-to)254-258
Number of pages5
JournalJournal of neurosurgery
Issue number2
StatePublished - 1990
Externally publishedYes


  • brain neoplasm
  • bromodeoxyuridine
  • cell kinetics
  • immunohistochemistry
  • iododeoxyuridine
  • monoclonal antibody

ASJC Scopus subject areas

  • Surgery
  • Clinical Neurology


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