目的研究人参总皂苷(TSPG)协同造血生长因子对CD34⁺细胞体外定向诱导分化为粒系血细胞的影响。方法应用阴性磁性分选策略,以StemsepTM系统从正常人骨髓细胞中分离CD34⁺造血干/祖细胞(HSC/HPC),通过甲基纤维素半固体培养法检测TSPG诱导CD34⁺HSC/HPC向粒系祖细胞集落(CFU-GM)增殖与分化的能力;在SCF+IL-3+GM-CSF+G-CSF(SIGG)液体培养体系中加入不同剂量的TSPG,检测对CD34⁺HSC/HPC增殖形成CFU-GM的影响以及CD33⁺细胞比例的变化。结果TSPG(10～50μg/mL)均能提高CD34⁺细胞形成CFU-GM的集落产率,以TSPG20μg/mL效果最为明显;不同质量浓度的TSPG协同造血生长因子在液体培养体系中诱导CD34⁺细胞2周,观察粒系血细胞的分化,TSPG10～70μg/mL均可不同程度地提高细胞总数、CFU-GM扩增倍数及CD34⁺细胞比例,TSPG20μg/mL是液体培养诱导CD34⁺细胞向粒系分化的最佳质量浓度。结论TSPG协同造血生长因子能促进CD34⁺细胞定向诱导分化为粒系血细胞。

Objective To investigate the effect of recombinant between hematopoietic growth factors and total saponins of Panax ginseng(TSPG)on granulocytopoietic differentiation of purified human CD34⁺ cells derived from human bone marrow cells.Methods A sterilized separation system for purification of CD34⁺ hematopoietic stem/progentior cells(HSC/HPC)was established by means of StemsepTM according to the strategy of negative selection on immunomagnetic separation.The capacity of various concentrations of TSPG inducing CD34⁺ cells to differentiate into CFU-GM cells was detected by methyl cellulose assay systems;CD34⁺ cells were committedly differentiated with various combination of cytokines in liquid culture systems and the expanded cell number,CD33⁺ cell ratio and granulocytopoietic progenitor cell number were detected;various concentrations of TSPG were then added into liquid culture systems containging SCF+IL-3+GM-CSF+G-CSF(SIGG),the elevation rate on proliferation of CD34⁺ cells and the formation of CFU-GM were observed and the ratio of CD33⁺ cells were estimated.Results TSPG(10—50 μg/mL)could increase the colony yield of CFU-GM,TSPG(20 μg/mL)did best.At the various concentrations of TSPG synergized with hematopoietic growth factors on granulocytopoietic differentiation of CD34⁺ cells for two weeks in liquid culture systems,TSPG(10—70 μg/mL)could increase the expanded cell number,the amplification multiple of total CFU-GM,and CD33⁺ cell ratio;TSPG 20 μg/mL was identified as the most potential concentration for the granulocytopoietic differentiation of CD34⁺ cells in liquid culture system.Conclusion TSPG may promote CD34⁺ cells to proliferate and differentiate to granulocytopoietic lineage by synergism with hematopoietic growth factors.

国家自然科学基金资助项目(编号38670880,30472253)