使用Percoll作为梯度介质对人单核白细胞进行不连续密度梯度分离。

Discontinuous density gradient separation of human mononuclear leucocytes using Percoll as gradient medium.

作者信息

Ulmer A J, Flad H D

出版信息

J Immunol Methods. 1979;30(1):1-10. doi: 10.1016/0022-1759(79)90268-0.

Abstract

The use of a new commerically available medium (Percoll) for fractionation of human mononuclear leucocytes is described. Cells can be fractionated on the basis of their densities with high reproducibility. The separated cells were characterized by morphological and functional criteria. Monocytes can be obtained in the low density fractions with a purity of 70%--90%. Lymphocytes were found in high density fractions with a purity up to 99%. No separation between E-rosette forming (E-RFC) and surface immunoglobulin-bearing lymphocytes was obtained. However, a reduced number of high avidity E-rosette forming lymphocytes (HAE-RFC) was found within low density lymphocytes. Best spontaneous DNA synthesis and reaction in mixed leucocyte culture (MLC) were obtained with cells isolated from the 1.066/1.068 density interface, whereas the stimulation with phytohemagglutinin (PHA) or pokeweed mitogen (PWM) had a peak response with cells from the 1.064/1.066 density interface. Colony forming myelopoietic stem cells and colony forming T-lymphocytes were detected in fractions of low density.

摘要

本文描述了一种新的市售介质（Percoll）用于人单核白细胞分级分离的方法。细胞可根据其密度进行分级分离，重现性高。通过形态学和功能标准对分离出的细胞进行了表征。单核细胞可在低密度级分中获得，纯度为70% - 90%。淋巴细胞存在于高密度级分中，纯度高达99%。未实现E花环形成细胞（E - RFC）和表面带有免疫球蛋白的淋巴细胞之间的分离。然而，在低密度淋巴细胞中发现高亲和力E花环形成淋巴细胞（HAE - RFC）数量减少。从密度为1.066/1.068的界面分离出的细胞在混合白细胞培养（MLC）中具有最佳的自发DNA合成和反应，而用植物血凝素（PHA）或商陆有丝分裂原（PWM）刺激时，密度为1.064/1.066的界面的细胞反应峰值最高。在低密度级分中检测到集落形成骨髓干细胞和集落形成T淋巴细胞。