Palutke M, KuKuruga D, Tabaczka P
Wayne State University School of Medicine, Department of Pathology, Berman Memorial Laboratories, Detroit, MI 48201.
J Immunol Methods. 1987 Dec 4;105(1):97-105. doi: 10.1016/0022-1759(87)90418-2.
A new method for measuring lymphocyte proliferation in response to mitogens and allogeneic cells without using radiolabelling is described. It utilizes flow cytometry and the monoclonal antibody, Ki-67, which detects a nuclear proliferation antigen. The entire test is performed in standard, 96-well tissue culture plates. Stable, clean nuclear suspensions rather than whole cells were used to avoid non-specific staining. The nuclei were stained by the indirect fluorescent method. Simultaneous measurements of DNA content were possible by dual staining with propidium iodide (PI). The percentage of Ki-67-positive nuclei correlated well with [3H]thymidine uptake and morphologic quantitation of blasts. This method avoids use of radioactive material and is less time consuming.
本文描述了一种无需使用放射性标记来测量淋巴细胞对有丝分裂原和同种异体细胞反应性增殖的新方法。该方法利用流式细胞术和单克隆抗体Ki-67，后者可检测一种核增殖抗原。整个检测在标准的96孔组织培养板中进行。为避免非特异性染色，使用的是稳定、纯净的核悬液而非全细胞。细胞核采用间接荧光法染色。通过与碘化丙啶（PI）进行双重染色可同时测量DNA含量。Ki-67阳性细胞核的百分比与[³H]胸腺嘧啶核苷摄取及母细胞的形态定量结果高度相关。该方法避免了放射性物质的使用，且耗时更短。
