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为探讨骨髓微环境中内皮细胞调节造血的可能机制 ,本实验研究了骨髓内皮细胞无血清条件培养液(BMEC CM )及其不同分子量组分对骨髓成纤维祖细胞 (CFU F)生长的作用。在体外培养条件下 ,收集人和小鼠纯骨髓内皮细胞无血清条件培养液 ,应用连续超滤技术从其中制备出分子量 >10kD ,3- 10kD和 <3kD三种组分 ,进行骨髓CFU F集落形成实验 ,检测这些条件培养液及其超滤组分的作用。结果显示 ,人和小鼠骨髓内皮细胞无血清条件培养液和分子量 <3kD超滤组分都抑制CFU F的生长 ,但是分子量 >10kD和 3- 10kD组分却不产生这种效应。两种条件培养液及其 <3kD组分不仅减少CFU F集落形成数 ,同时也使集落变小 ,而且 ,<3kD组分与CFU F集落数之间有明显的负相关浓度依赖性关系。这些结果提示 ,骨髓内皮细胞至少能分泌一种分子量小于 3kD的对骨髓CFU F生长有抑制作用的细胞因子。
In order to explore the possible mechanism of hematopoietic regulation of endothelial cells in bone marrow microenvironment, we studied the effects of BMEC CM and different molecular weight components on the growth of bone marrow fibroblast progenitor cells (CFU F). Under in vitro culture conditions, serum-free conditioned medium of pure bone marrow endothelial cells of human and mouse were collected and three components of molecular weight> 10kD, 3-10kD and <3kD were prepared by continuous ultrafiltration technique, and bone marrow CFU F colonies Experiments were performed to examine the effect of these conditioned media and their ultrafiltration components. The results showed that both serum-free conditioned medium of human and mouse bone marrow endothelial cells and the ultrafiltration fraction of <3 kD in molecular weight inhibited the growth of CFU F, but did not produce this effect with components of> 10 kD and 3 to 10 kD. Both conditioned medium and its <3 kD fraction not only decreased CFU F colony formation but also decreased colonies, and there was a significant negative concentration-dependent relationship between <3 kD fraction and CFU F colonies. These results suggest that bone marrow endothelial cells secrete at least one cytokine that inhibits bone marrow CFU F growth with a molecular weight less than 3 kD.