van den Akker Erica L T, Baan Carla C, van den Berg Bart, Russcher Henk, Joosten Koen, Hokken-Koelega Anita C S, Lamberts Steven W J, Koper Jan W
Department of Pediatric Endocrinology, Office sp 1536, Erasmus MC, PO Box 2060, 3000 CB Rotterdam, The Netherlands.
Intensive Care Med. 2008 May;34(5):912-6. doi: 10.1007/s00134-007-0989-0. Epub 2008 Jan 8.
To determine the cell content and purity of Ficoll-separated peripheral blood mononuclear cells and granulocyte isolates in sepsis patients compared to healthy controls.
Prospective study in the adult and pediatric intensive care departments of the Erasmus University Medical Center in the Netherlands.
Three sepsis patients (two adults, one child) and four healthy controls.
Blood leukocytes were separated by Ficoll into an interface and a bottom fraction. The cell content and purity was analyzed by cytospin and flow-cytometric immunofluorescence. In sepsis patients, the interface consisted of 11-52% mononuclear cells only, due to high contamination with granulocytes (48-89%). This was in contrast to a high proportion of mononuclear cells (88-100%) in healthy controls. The bottom fraction showed a cell purity of >or=92% polymorphonuclear granulocytes in sepsis patients as well as in healthy controls.
Ficoll-separated leukocytes of sepsis patients are not suitable for studying mononuclear cells but can be used for studying granulocytes with high purity. The mononuclear cell fraction is highly contaminated with granulocytes. Additional separation techniques are necessary to obtain a pure cell fraction.
与健康对照相比，确定脓毒症患者中经Ficoll分离的外周血单个核细胞和粒细胞分离物的细胞含量及纯度。
在荷兰伊拉斯姆斯大学医学中心的成人及儿科重症监护病房进行的前瞻性研究。
3例脓毒症患者（2例成人，1例儿童）和4例健康对照。
通过Ficoll将血液白细胞分离为界面层和底层部分。通过细胞离心涂片法和流式细胞免疫荧光法分析细胞含量及纯度。在脓毒症患者中，由于粒细胞污染严重（48%-89%），界面层仅由11%-52%的单个核细胞组成。这与健康对照中高比例的单个核细胞（88%-100%）形成对比。在脓毒症患者及健康对照中，底层部分显示多形核粒细胞的细胞纯度≥92%。
脓毒症患者经Ficoll分离的白细胞不适合用于研究单个核细胞，但可用于高纯度粒细胞的研究。单个核细胞部分被粒细胞高度污染。需要额外的分离技术来获得纯细胞部分。
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