一种使培养中的正常人类细胞和恶性人类细胞同步化的方法。

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

作者信息

Hill B T, Whelan R D, Hemmings V J, Franks L M

出版信息

Cell Biol Int Rep. 1977 Jul;1(4):379-84. doi: 10.1016/0309-1651(77)90069-8.

Abstract

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.

摘要

描述了一种在正常间充质和转化上皮细胞中实现可重复的S期准同步化的方法。将细胞以低密度接种于含10%血清的培养基中。24小时后，将血清浓度降至0.5%。110小时后，在含10%血清加2.5mM羟基脲的新鲜培养基中，于G1/S边界收集细胞20小时。去除羟基脲并进行胰蛋白酶消化后，重新接种的细胞高度同步地进入S期，这通过放射自显影、用3H-胸腺嘧啶脉冲标记、细胞生长和延时摄影来判断。同步化后6小时，80%的细胞群体进入S期，10 - 13小时之间70%的细胞经历有丝分裂。