Dettman G L, Wilbur S M
J Immunol Methods. 1979;27(3):205-17. doi: 10.1016/0022-1759(79)90218-7.
A method is described for the inexpensive and easy preparation of colloidal silica--aluminum modified--polyvinylpyrrolidone (CS-AM-PVP) density gradient centrifugation medium. Using density gradient centrifugation, several cell separation and biochemical characteristics were studied: centrifuge tube wall cell adherence, mouse spleen cell density distribution, rebanding properties, mitogen response and cell aggregation. Cell adherence to the centrifuge tube wall using density gradients of CS-AM-PVP was compared with density gradients of bovine serum albumin and Ficoll. Few cells adhered to the centrifuge tube wall when CS-AM-PVP was used as a gradient medium; whereas, significant cell adherence to the centrifuge tube cell wall occurred when bovine serum albumin or Ficoll was used as a gradient medium. The CS-AM-PVP gradient medium did not inhibit the response to mitogens of mouse spleen cells which had been separated into density subpopulations in a discontinuous CS-AM-PVP density gradient, caused a minimum amount of cell aggregation, and was found to be non-toxic.
描述了一种制备胶体二氧化硅 - 铝改性 - 聚乙烯吡咯烷酮(CS - AM - PVP)密度梯度离心介质的方法,该方法成本低廉且操作简便。利用密度梯度离心,研究了几个细胞分离和生化特性:离心管壁细胞黏附、小鼠脾细胞密度分布、再分层特性、有丝分裂原反应和细胞聚集。将使用CS - AM - PVP密度梯度时细胞对离心管壁的黏附情况与使用牛血清白蛋白和聚蔗糖密度梯度时的情况进行了比较。当使用CS - AM - PVP作为梯度介质时,很少有细胞黏附到离心管壁;而当使用牛血清白蛋白或聚蔗糖作为梯度介质时,会有大量细胞黏附到离心管壁。CS - AM - PVP梯度介质不会抑制在不连续CS - AM - PVP密度梯度中已分离成密度亚群的小鼠脾细胞对有丝分裂原的反应,引起的细胞聚集量最少,且无毒。