Synchronization of cancer cell lines with methotrexate in vitro

Eugenio Erba; Soumitra Sen

doi:10.1007/BF00122166

Synchronization of cancer cell lines with methotrexate in vitro

Eugenio Erba, Soumitra Sen

Istituto di Ricerche Farmacologiche "Mario Negri"

Research output: Contribution to journal › Article › peer-review

Abstract

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 language	English
Pages (from-to)	149-163
Number of pages	15
Journal	Methods in Cell Science
Volume	18
Issue number	2
DOIs	https://doi.org/10.1007/BF00122166
Publication status	Published - 1996

Keywords

Cancer cell lines
Cell synchronization
Flow cytometry
MTX

ASJC Scopus subject areas

Cell Biology

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10.1007/BF00122166

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AU - Sen, Soumitra

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AB - 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.

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Synchronization of cancer cell lines with methotrexate in vitro

Abstract

Keywords

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