Isolation of neutrophil precursors from bone marrow for biochemical and transcriptional analysis
Isolation of neutrophil precursors from bone marrow for biochemical and transcriptional analysis
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DOI：
10.1016/s0022-1759(99)00176-3
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发表时间：
1999-12-17
影响因子：
2.2
通讯作者：
Borregaard, N
中科院分区：
文献类型：
--
作者：
Cowland, JB;Borregaard, N
嗜中性粒细胞是外周血中数量最多的白细胞。从多能祖细胞到成熟中性粒细胞的发育在骨髓中发生10-14天。为了了解这一过程背后的细胞机制，有必要研究来自中性粒细胞分化不同阶段的细胞。由于没有人细胞系能够忠实地再现从早幼粒细胞到分叶嗜中性粒细胞的整个分化过程，因此必须对来自人骨髓的嗜中性粒细胞前体进行许多成熟依赖性过程的分析。为此目的，需要一种技术，其中中性粒细胞前体可以从骨髓中分离，并根据其成熟度进行分离。两种不同的方法已被证明是有用的未成熟的中性粒细胞的分离：密度离心Percoll梯度，其中的细胞密度增加与成熟形成的分离的基础，和多维流式细胞术，是一个组合的大小，颗粒，和表面标记用于区分不同的中性粒细胞前体。本文将回顾这两种分离中性粒细胞前体的方法，特别强调Percoll密度离心以及使用该技术分离的细胞分析中性粒细胞特异性mRNA和中性粒细胞颗粒蛋白的生物合成。(C)1999 Elsevier Science B. V.保留所有权利。
The neutrophilic granulocyte is the most numerous leukocyte in peripheral blood. The development from a multipotent progenitor cell to a mature neutrophil takes place in the bone marrow over a period of 10-14 days. In order to understand the cellular mechanisms behind this process, it is necessary to investigate cells from different stages of neutrophil differentiation. As no human cell line has the ability to faithfully reproduce the entire differentiation process from promyelocyte to segmented neutrophil the analysis of many maturation-dependent processes has to be done on neutrophil precursors from human bone marrow. For this purpose, a technique whereby neutrophil precursors can be isolated from the bone marrow and separated according to their maturity is required. Two different methods have been shown to be useful for isolation of immature neutrophils: density centrifugation on a Percoll gradient, where the increasing density of the cells with maturity forms the basis of the separation, and multidimensional flow cytometry, were a combination of size, granulation, and surface makers are used for the discrimination of different neutrophil precursors. This paper will review these two methods for separation of neutrophil precursors with special emphasis on Percoll density centrifugation and the use of cells isolated by this technique for the analysis of neutrophil-specific mRNAs and the biosynthesis of neutrophil granule proteins. (C) 1999 Elsevier Science B.V. All rights reserved.