小鼠骨髓内皮细胞无血清条件培养液及其超滤组分对粒巨噬系祖细胞生长的影响

EFFECTS OF BONE MARROW ENDOTHELIAL CELL CONDITIONED MEDIUM AND ITS ULTRAFILTRATION PREPARED COMPONENTS ON THE GROWTH OF CFU GM *

本研究通过传代培养小鼠骨髓内皮细胞系细胞,收集无血清条件培养液(mBMECCM),将其作多级串联超滤,获得分子量大于10、3～10、1～3、05～1kD和小于05kD超滤组分。进行粒巨噬系造血祖细胞集落形成试验,检测了它们的作用。mBMECCM和3～10kD组分对粒巨噬系祖细胞(CFUGM)生长未见明显影响,而分子量大于10kD和05～1kD组分促进CFUGM的增殖,分子量1～3kD和小于05kD组分则抑制CFUGM的增殖。这4种超滤组分对CFUGM生长的效应均有剂量依赖性。这些结果提示,在体外培养条件下,小鼠骨髓内皮细胞分泌若干种活性成分。

In this study, we obtained the serum free conditioned medium from subcultures of murine bone marrow derived endothelial cell line which has been established recently by ourselves. And then, molecular weight (MW) >10 kD, 3～10 kD, 1～3 kD, 0 5～1 kD and <0 5 kD components were sifted out from the conditioned medium (mBMEC CM) by means of serial ultrafiltration. Assays of granulocyte macrophage colony forming cell (CFU GM) were performed to examine the effects of mBMEC CM and its ultrafiltration prepared components. It was not observed that mBMEC CM and MW 3～10 kD component had any significant influence on the growth of CFU GM. However, MW>10 kD and 0 5～1 kD components enhanced the proliferation of CFU GM, whereas MW 1～3 kD and <0 5 kD ones inhibited it. All these four components exerted their effects on the growth of CFU GM in dose dependent way. Our observations suggest that under the conditions of in vitro, murine bone marrow endothelial cells produce several active components which have promoting or inhibitory effects on the growth of CFU GM.