A method of synchronization of normal and malignant human cells in culture

A method is described for providing reproducible S phase parasynchrony in both normal mesenchyme and transformed epithelia. Cells were seeded at low density in medium containing 10% serum. 24 h later the serum concentration was reduced to 0.5%. After 110th the cells were collected at the G1/S boundary in fresh medium containing 10% serum plus 2.5mM hydroxyurea over 20h. After removal of hydroxyurea and trypsinization the re-plated cells entered the S phase with a high degree of synchrony, as judged by autoradiography, pulse-labelling with 3H-thymidine, cell growth and time lapse cinematography. By 6h after synchronization 80% of the population had entered the S phase and between 10-13h 70% went through mitosis.