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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。
The synthesis, accumulation and correlation of DNA, RNA and PRO are the three most important parameters that regulate the cell cycle kinetics. Simultaneous detection of these components allows more precise and detailed assessment of the cell cycle kinetic characteristics. In this paper, the content of macrophages DNA, RNA and PRO in human normal bone marrow CD34 ~ + hematopoietic cells was explored in order to understand the characteristics of the cycle kinetics of CD34 ~ + hematopoietic cells. The new type of CIMS-100 immunomagnetic separation system was used to efficiently enrich human bone marrow CD34 + hematopoietic cells. The purity of CD34 + hematopoietic cells enriched from 90 to 95% was identified by FCM and APAAP. Subsequently, DNA, RNA and PRO were labeled with propidium iodide (PI), Pyronin Y (PY) and fluorescein isothiocyanate (FITC) respectively and detected by FCM. The results showed that the content of DNA, RNA and PRO in CD34 ~ + hematopoietic cells was significantly lower than that of mononuclear cells, accounting for 34 ± 3%, 48 ± 21% and 62 ± 14% of the latter respectively. Combined with our previous results, we believe that CD34 ~ + hematopoietic cells are indeed a unique somatic cell population, not only in the reconstruction of hematopoietic and immunological functions, but also in the cell cycle kinetics. To the best of our knowledge, this is the first systematic review of the molecular dynamics of human CD34 + hematopoietic cell cycle in the world and provides direct evidence that most CD34 + hematopoietic cells are in quiescence.