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在体内,正常的造血过程是通过骨髓造血微环境来维持的.研究表明,这种微环境是一个复杂的三维结构.由骨髓基质细胞如纤维母细胞、血管内皮细胞、巨噬细胞、脂肪细胞等组成.这些基质细胞能分泌造血细胞因子,并能维持造血细胞在体外长期生存.根据这一原理,我们建立了一种新颖的小鼠HPC培养系统.该系统以骨髓微血管内皮细胞(BMEC)作为生长支持介质,通过加入重组人粒细胞集落刺激因子(G-CSF)来促进HPC增殖.实验结果表明,将氟脲嘧啶(5-Fu,150mg/kg) 处理的C57小鼠骨髓细胞置于BMEC细胞层上,并加入25ng/ml G-CSF培养2周,细胞数量增加21.2倍.流式细胞分析提示,扩增后的细胞中60%表达祖细胞相关抗原CD117,40%表达髓系细胞分化抗原Gr-1.而培养前的骨髓细胞中上述二种细胞的比例分别为5%和
In vivo, the normal hematopoietic process is maintained by the bone marrow hematopoietic microenvironment. Studies have shown that this microenvironment is a complex three-dimensional structure. It consists of bone marrow stromal cells such as fibroblasts, vascular endothelial cells, macrophages, and adipocytes. Such components. These stromal cells can secrete hematopoietic cytokines and can maintain long-term survival of hematopoietic cells in vitro. Based on this principle, we have established a novel mouse HPC culture system. The system uses bone marrow microvascular endothelial cells (BMEC). As a growth support medium, HPC proliferation was promoted by adding recombinant human granulocyte colony-stimulating factor (G-CSF). Experimental results showed that the C57 mouse bone marrow cells treated with fluorouracil (5-Fu, 150 mg/kg) were placed On the BMEC cell layer, 25 ng/ml G-CSF was added and cultured for 2 weeks. The number of cells increased by 21.2 fold. Flow cytometry analysis showed that 60% of the amplified cells express CD117 and 40% of myeloid cells. Differentiating the antigen Gr-1. The percentage of the above two kinds of cells in the bone marrow cells before culture was respectively 5% and