人造血干/祖细胞多态性的研究:Ⅺ.人正常骨髓CD34^+造血细胞周期动力学相关大分子DNA、RNA和PRO含量的分析  

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：奚永志[1] 张志欣[2] 张双喜[3] 孔繁华[1] 李秀森[3] 唐佩弦[3] 魏文[3] 金荔[1] 陈兴国[1] 

机构地区：[1]军事医学科学院附属医院免疫学研究室,北京100039 [2]北京市血液中心白细胞实验室,北京100010 [3]军事医学科学院基础医学研究所,北京100850

出　　处：《实验生物学报》1998年第2期129-135,共7页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金~~

摘　　要：DNA、RNA和PRO的合成、积累及相互关系是调控细胞周期动力学最主要的三个参数。同时检测这些组成部分能够更精确细致地评判细胞的周期动力学特征。本文探索了人正常骨髓CD34^+造血细胞周期动力学相关大分子DNA、RNA和PRO的含量,以便认识CD34^+造血细胞周期动力学的特征。用新型CIMS-100免疫磁性分离系统高效富集人骨髓CD34^+造血细胞,经FCM及APAAP鉴定,富集的CD34^+造血细胞的纯度达90～95%。随之采用碘化丙啶(PI)、派若宁Y(PY)及异硫氰荧光素(FITC)分别进行标记DNA、RNA和PRO并在FCM上检测。结果表明,DNA、RNA和PRO在CD34^+造血细胞中的含量明显低于单个核细胞,分别仅占后者的34±3%、48±21%及62±14%。结合我们以往的结果,我们认为CD34^+造血细胞的确是一独特的体细胞群,不仅表现在重建造血与免疫学功能上,而且表现在细胞周期动力学上。据我们所知,这是目前国际上首次有关人CD34^+造血细胞周期动力学相关大分子的系统分析报道,提供了大多数CD34^+造血细胞处于静止期的直接证据。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 pro-pidium iodide, pyronin Y and fluorescein isothio-cyante 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.

关 键 词：人 正常骨髓 造血细胞 周期动力学 DNA PRO 

分 类 号：R331.2[医药卫生—人体生理学]