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[人类造血干细胞/祖细胞的多样性:IX. 人骨髓CD34+造血细胞中与细胞周期动力学相关的大分子DNA、RNA和蛋白质含量]

[The diversity of human hematopoietic stem/progenitor cells: IX. Cell-cycles kinetics related macromolecules DNA, RNA and protein contents in CD34+ hematopoietic cells of human bone marrow].

作者信息

Xi Y Z, Zhang Z X, Zhang S X, Kong F H, Li X S, Tang P H, Wei W, Jin L, Chen X G

机构信息

Department of Transplantation Immunology, Affiliated 307 Hospital for Academy of Medical Sciences, Beijing 100039.

出版信息

Shi Yan Sheng Wu Xue Bao. 1998 Jun;31(2):129-35.

Abstract

DNA, RNA and PRO comprise the bulk of macromolecules in cells, which have been proven to play an important role in regulating cell cycle transverse capacity, cell division, growth, and size. Simultaneous analysis of these moieties could provide more comprehensive and accurate information on cell cycle kinetics. In this study, DNA, RNA and PRO contents related to cell cycle kinetics in CD34+ hematopoietic cells of human bone marrow were measured to understanding the cell cycle kinetic features in CD34+ hematopoietic cells. For this reason, CIMS-100 immunomagnetic isolator, a novel isolation system, was used to enrich efficiently CD34+ hematopoietic cells from human bone marrow. The purity of enriched CD34+ hematopoietic cells determined by both FACS and APAAP staining is ranging from 90%-95%. Cellular DNA, RNA and PRO were stained with fluorochromes propidium iodide, pyronin Y and fluorescein isothiocyante respectively The fluorescence intensities reflecting the DNA, RNA and PRO content of individual cell were analyzed in FACS can by different excitation wavelengthes. DNA, RNA and PRO contents in CD34+ hematopoietic cells were far lower than these of bone marrow mononuclear cells, only being 34 +/- 3% (DNA), 48 +/- 21% (RNA) and 62 +/- 14% (PRO) of BMMNCs respectively. Collectively, these data combined with previous results from both ours and others indicated that CD34+ hematopoietic cells are indeed an unique cell population, not only in reconstitute of hematopoietic and immunological functions, but also in cell cycle kinetics. This is, to our knowledge, the first detailed report on the analysis of DNA, RNA and PRO contents related to cell cycle kinetics in CD34+ hematopoietic cells. And the results provide more direct evidence that the majority of CD34+ hematopoietic cells are in resting state.

摘要

DNA、RNA和蛋白质构成了细胞中大部分的大分子,已被证明在调节细胞周期转换能力、细胞分裂、生长和大小方面发挥重要作用。对这些成分的同时分析可以提供关于细胞周期动力学更全面和准确的信息。在本研究中,测量了人骨髓CD34+造血细胞中与细胞周期动力学相关的DNA、RNA和蛋白质含量,以了解CD34+造血细胞的细胞周期动力学特征。因此,一种新型分离系统CIMS-100免疫磁珠分离器被用于从人骨髓中高效富集CD34+造血细胞。通过流式细胞术(FACS)和碱性磷酸酶抗碱性磷酸酶(APAAP)染色测定的富集CD34+造血细胞的纯度在90%-95%之间。细胞DNA、RNA和蛋白质分别用荧光染料碘化丙啶、派洛宁Y和异硫氰酸荧光素进行染色。在流式细胞仪中通过不同的激发波长分析反映单个细胞DNA、RNA和蛋白质含量的荧光强度。CD34+造血细胞中的DNA、RNA和蛋白质含量远低于骨髓单个核细胞,分别仅为骨髓单个核细胞的34±3%(DNA)、48±21%(RNA)和62±14%(蛋白质)。总体而言,这些数据与我们和其他人之前的结果表明,CD34+造血细胞确实是一个独特的细胞群体,不仅在造血和免疫功能重建方面,而且在细胞周期动力学方面。据我们所知,这是关于CD34+造血细胞中与细胞周期动力学相关的DNA、RNA和蛋白质含量分析的第一份详细报告。结果提供了更直接的证据,表明大多数CD34+造血细胞处于静止状态。

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