Kriegler A B, Bradley T R, Hodgson G S, McNiece I K
Cell Biology Group, Peter MacCallum Cancer Institute, Melbourne, Victoria, Australia.
J Immunol Methods. 1987 Oct 23;103(1):93-102. doi: 10.1016/0022-1759(87)90246-8.
Synergistic factors from media conditioned (CM) by human placentas or the 5637 human bladder carcinoma cell line (SFH-HPCM and SFH-5637 respectively) have the ability to stimulate early progenitor cells in mouse bone marrow to form large colonies in agar cultures after 12-14 days, in the presence of CSF-1. Culture conditions have been examined and a quicker and more convenient liquid culture assay has been developed for this factor, using a tetrazolium salt to quantitate cell proliferation. The use of flat-bottomed vessels, high cell density, supra-optimal doses of CSF-1 or the addition of WEHI-3-CM to these cultures, all resulted in a decrease in the required incubation time. In combination, these modifications reduced the assay time to 4 days.
来自人胎盘条件培养基(CM)或5637人膀胱癌细胞系(分别为SFH - HPCM和SFH - 5637)的协同因子,在存在集落刺激因子-1(CSF - 1)的情况下,有能力刺激小鼠骨髓中的早期祖细胞在琼脂培养中12 - 14天后形成大的集落。已经研究了培养条件,并开发了一种更快、更方便的液体培养测定法来检测该因子,使用四氮唑盐来定量细胞增殖。使用平底容器、高细胞密度、超最佳剂量的CSF - 1或向这些培养物中添加WEHI - 3 - CM,所有这些都导致所需的孵育时间减少。综合起来,这些改进将测定时间缩短至4天。