Synchronization of cancer cell lines with methotrexate in vitro

Eugenio Erba, Soumitra Sen

Research output: Contribution to journalArticlepeer-review


Detailed procedure are described for inducing synchrony in cancer cell lines of human origin (Jurkat, K562, U937, SW626) and L1210 cell line of murine origin by using very low non-toxic concentrations (0.04-0.08 μM for 13-24 hours) of methotrexate under standard culture conditions. This protocol offers a method for synchronization of cells at the G1/S boundary and through the S-G2-M phases of the cell cycle. Kinetic behavior and biological properties of the synchronized cells are considered for characterisation of the system. Comparisons are made with MTX synchronization and that induced by aphidicolin alone or by a combination of serum deprivation and aphidicolin. Methotrexate appears to be the best choice for obtaining highly synchronous human cancer cell population without cytotoxicity or detectable physiological perturbations.

Original languageEnglish
Pages (from-to)149-163
Number of pages15
JournalMethods in Cell Science
Issue number2
Publication statusPublished - 1996


  • Cancer cell lines
  • Cell synchronization
  • Flow cytometry
  • MTX

ASJC Scopus subject areas

  • Cell Biology


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