PUCK T T, CIECIURA S J, ROBINSON A
J Exp Med. 1958 Dec 1;108(6):945-56. doi: 10.1084/jem.108.6.945.
A methodology designed to eliminate mitotic inhibitor action and involving use of pretested fetal calf serum and careful pH and temperature control has been described by which cells from normal human and animal tissue can be maintained in active growth for long periods in vitro without development of aneuploidy. By means of this procedure, it is possible reliably to establish cell cultures from minute skin biopsies which can be taken from any individual. Clones of mammalian cells with chromosomal markers have been isolated by this means from x-irradiated non-irradiated cell cultures. Application of these techniques to chromosome delineation in large numbers of human subjects; determination of chromosomal sex in patients; spontaneuos and induced genetic changes in somatic mammalian cells in vivo and in vitro; comparison of metabolic differences between normal and cancerous cells and other problems have been indicated.
已经描述了一种旨在消除有丝分裂抑制剂作用的方法,该方法涉及使用预先检测过的胎牛血清以及仔细的pH值和温度控制,通过这种方法,来自正常人和动物组织的细胞可以在体外长期保持活跃生长而不会出现非整倍体。通过这个程序,可以可靠地从任何个体的微小皮肤活检样本中建立细胞培养物。通过这种方法,已经从经X射线照射和未经照射的细胞培养物中分离出具有染色体标记的哺乳动物细胞克隆。这些技术在大量人类受试者的染色体描绘中的应用;患者染色体性别的确定;体内和体外体细胞哺乳动物细胞的自发和诱导遗传变化;正常细胞和癌细胞之间代谢差异的比较以及其他问题已经得到了说明。