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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。  相似文献   
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人CD34~ 造血细胞是具有高度自我更新、多向分化及重建长期造血与免疫学功能的独特体细胞。为系统探索CD34~ 造血细胞的形态、细胞化学及超微结构特征,新近我们设计组合并建立了CIMS-100-FACS 440无菌二次分选术,可使所获CD34~ 造血细胞的纯度达100%。在此基础上,本研究采用Cambri-dge Quantimet 970全自动图像分析仪对光学显微镜、扫描电镜及透射电镜下的CD34~ 造血细胞进行了体视学方面的某些探讨,进一步从三维结构信息中深刻揭示CD34~ 造血细胞的形态计量学特征。经扫描→模数转换←阴影校正→图像暂存←统计分析等检测,结果表明:CD34~ 造血细胞的直径3.490—6.741μm,周长11.776—26.240μm,面积9.565—35.686μm~2,形状因子1.048—1.840,核浆比0.58—0.72,平均光密度0.17675—0.65100,积分光密度2717.217—9870.643。由此可见CD34~ 造血细胞的确为非均一细胞群,这可能与CD34~ 造血细胞的功能亚群与分化阶段密切相关。据我们所知,这是国际上首次有关人CD34~ 造血细胞体视学特征的报道。  相似文献   
3.
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.  相似文献   
4.
人CD34+造血细胞是具有高度自我更新,多向分化及重建长期造血与免疫学功能的独特体细胞。为系统探索CD34+造血细胞的形态,细胞化学及超微结构特征,新近我们设计组合并建立了CIMS-100FACS440无菌二次分选术,可使所获CD34+5造血的纯度达100%。  相似文献   
5.
mPEG修饰HLA抗原的方法学研究   总被引:2,自引:0,他引:2  
为了研究mPEG有效修饰HLA抗原,阻断HLA抗原与其相应抗体间特异性免疫反应的方法.在22℃,pH7.4的PBS介质中,采用浓度梯度法修饰淋巴细胞表面HLA抗原.结果显示经mPEG处理后的淋巴细胞表面HLA-1类抗原与其相应抗体间的微量淋巴细胞毒试验为阴性.由此可以得出结论,经mPEG修饰后可以完全阻断HLA—I类抗原与其相应抗体间的特异性免疫反应.  相似文献   
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