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使用新型血细胞分离器制备的血沉棕黄层中单核细胞的快速分离

Rapid isolation of mononuclear cells from buffy coats prepared by a new blood cell separator.

作者信息

Figdor C G, Bont W S, De Vries J E

出版信息

J Immunol Methods. 1982 Dec 17;55(2):221-9. doi: 10.1016/0022-1759(82)90034-5.

Abstract

A newly developed cell separator for the preparation and fractionation of buffy coat cells from human peripheral blood is described. In this cell separator buffy coats (BC-1) as routinely obtained from blood banks were used for the preparation of a second buffy coat (BC-2) with a volume of only 5-6 ml. A special fractionation device allowed sterile isolation of almost pure platelets and mononuclear cells with recoveries of 75 +/- 10% and 89 +/- 4% respectively. The white blood cell contamination of the platelet suspension never exceeded 20 X 10(6) leukocytes (i.e., less than 1 leukocyte per 5000 platelets). Furthermore, the mononuclear cell suspensions were shown to be contaminated with only 3 +/- 2% granulocytes, whereas the white blood cell/red blood cell ratio was 2.6 +/- 1.6, so that they could therefore be directly used for further separation by means of centrifugal elutriation. These results indicate that this cell separator provides a rapid (+/- 1 h) isolation of both platelets and mononuclear cells without exposing the buffy coat cells to foreign substances like Ficoll or Percoll.

摘要

本文描述了一种新开发的用于从人外周血中制备和分离血沉棕黄层细胞的细胞分离器。在这种细胞分离器中,使用从血库常规获得的血沉棕黄层(BC-1)来制备体积仅为5-6毫升的第二种血沉棕黄层(BC-2)。一种特殊的分离装置可无菌分离出几乎纯的血小板和单核细胞,回收率分别为75±10%和89±4%。血小板悬液中的白细胞污染从未超过20×10⁶个白细胞(即每5000个血小板中白细胞少于1个)。此外,单核细胞悬液显示仅被3±2%的粒细胞污染,而白细胞/红细胞比例为2.6±1.6,因此它们可直接用于通过离心淘析进行进一步分离。这些结果表明,这种细胞分离器能够快速(±1小时)分离血小板和单核细胞,而不会使血沉棕黄层细胞接触到诸如聚蔗糖或 Percoll 等外来物质。

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