内皮细胞对小鼠粒系、红系和巨核系造血的调控作用

Effect of Human Umbilical Vein Endothelial Cells on Granulopoiesis, E rythropoiesis and Megakaryopoiesis in Mice

目的 了解内皮细胞对小鼠粒系、红系和巨核系造血功能的影响 ,为进一步研究血发生及其调控提供理论依据。方法 取剖腹产术后 12小时以内的人脐静脉内皮细胞 ,按 Jaffe法进行培养 ,建立内皮细胞体外培养模型。而后收集原代培养的内皮细胞上清液 ,应用造血祖细胞体外培养方法进行体外粒 -单系祖细胞克隆形成单位 ( CFU - GM)、早期红系祖细胞克隆形成单位 ( BFU - E)、晚期红系祖细胞克隆形成单位 ( CFU - E)和巨核系祖细胞克隆形成单位 ( CFU - MK)半固体培养。结果 在有内皮细胞上清液存在的各培养体系中 ,集落形成良好。而在同样培养条件下 ,在无内皮细胞上清液存在的体系中 ,需加入外源性的 GM- CSF、EPO和 IL - 3才能表现出对粒系、红系和巨核系造血的刺激作用。结论 1内皮细胞能够通过分泌造血生长因子 ,支持各系祖细胞的增殖与分化 ;2内皮细胞上清液可作为标准的生长因子来源 ,用于体外干细胞的扩增和某些血液病的治疗 ;3内皮细胞和造血干细胞 /祖细胞的相互作用在体内的造血调控中也可能起了重要作用 ;4在内皮细胞上清液中可能含有尚未能证明的其他造血生长因子。这些因子单独或协同地发挥作用 。

Objective To investigate the effects of endothe lial cells on the p roliferations of granulocytic, erythrocytic and megakaryocytic progenitor cells. Methods Human umbilical vein endothelial cells were collected within 12 hours a fter cesarean section and were cultured according to Jaffe's method. The human e ndothelial culture supernatant (HECS) harvested from primary culture medium was centrifuged. Bone marrow mononuclear cells of mice were prepared and then cultur ed in semi solid agar or methylcellulose with the addition of HECS. Res ults Colo ny forming yields were increased in the culture systems with HECS. But in the s y stems with no HECS, exogenous GM CSF,EPO and IL 3 added to the hematopoietic progeni tor cultures were necessary to induce colony forming. Synergistic increase in p r ogenitor cell expansion was observed when both HECS and exogenous cytokines wer e added to hematopoietic progenitor cultures. Conclusion ①End othelial cells ca n support the proliferation and differentiation of hematopoietic stem and progen itor cells. ②HECS as a source of standard hematopoietic growth factors can be u sed in the expansion of stem cells in vitro and in the therapy of hemopathy. ③I nteractions between endothelial cells and hematopoietic stem cell or progenitors may play an important role in the regulation of hematopoiesis in vivo. ④Endoth elial cells may produce other soluble growth factors potentiating the action of a set of cytokines such as GM CSF and IL 6.