Rapid determination of DNA synthesis in adherent cells grown in microtiter plates

Min Kyung H Song, Henry Krutzsch, W. David Hankins, William L. Richards, Snorri S. Thorgeirsson

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

A specific, rapid, and economical method for measuring the extent of DNA synthesis in adherent rat hepatoma H4-II-E cells grown in 96-well microtiter plates is described. The adherent cells were pulsed for 1 h with [methyl-3H]thymidine, released from the substratum by trypsinization, and collected on fiberglass filters with a MASH II cell harvester. The amount of radioactivity incorporated was directly proportional to the number of cells per well. Growth curves generated by measuring [methyl-3H]thymidine incorporation and counting the number of cells per well were identical. Experiments with inhibitors of DNA, protein, and RNA synthesis demonstrated that this method selectively measured DNA synthesis. In addition, [3H]thymidine uptake showed excellent correlation with autoradiographic assessment of DNA synthesis. This specific and sensitive method for determining DNA synthesis in microtiter cultures should facilitate studies of effects of various growth-controlling agents on epithelial, fibroblastic, and other cells which grow as adherent cells in culture.

Original languageEnglish (US)
Pages (from-to)271-276
Number of pages6
JournalExperimental Cell Research
Volume156
Issue number1
DOIs
StatePublished - 1985
Externally publishedYes

ASJC Scopus subject areas

  • Cell Biology

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