Ma Li-xia, Huang Yan-hong, Cheng La-mei, Lei Jun, Wang Qi-ru
Experimental Hematology Laboratory, Department of Physiology, Xiangya Medical School of Central South University, Changsha 410078, China.
Chin Med J (Engl). 2007 Jul 5;120(13):1192-7.
Macrophage stimulating protein (MSP) is produced by human bone marrow endothelial cells. In this study, we sought to observe its effects on inducing the expansion of early hematopoietic progenitor cells which were cultured in a liquid culture system in the presence of the combination of stem cell factor (SCF), interleukin 3 (IL-3), interleukin 6 (IL-6), granulocyte macrophage-colony stimulating factor (GM-CSF), erythropoietin (EPO) (Cys) and MSP or of Cys and bone marrow endothelial cell conditioned medium (EC-CM).
Human bone marrow CD34(+) cells were separated and cultured in a liquid culture system for 6 days. Granulocyte-macrophage colony forming unit (CFU-GM) and colony forming unit-granulocyte, erythrocyte, macrophage, megakaryocyte (CFU-GEMM) were employed to assay the effects of different treatment on the proliferation of hematopoeitic stem/progenitor cells. The nitroblue tetrazolium (NBT) reductive test and hoechest 33258 staining were employed to reflect the differentiation and apoptosis of the cells respectively.
MSP inhibited the proliferation of CFU-GM and CFU-GEMM in semi-solid culture and the inhibitory effect on CFU-GEMM was stronger than on CFU-GM. MSP inhibited the differentiation of early hematopoietic progenitor cells induced by hematopoietic stimulators. Bone marrow (BM) CFU-GEMM was 2.3-fold or 1.7-fold increase or significantly decreased in either Cys + EC-CM, Cys + MSP or Cys compared with 0 hour control in liquid culture system after 6 days.
MSP, a hematopoietic inhibitor, inhibits the differentiation of early hematopoietic progenitor cells induced by hematopoietic stimulators and makes the early hematopoietic progenitor cells expand in a liquid culture system.
巨噬细胞刺激蛋白（MSP）由人骨髓内皮细胞产生。在本研究中，我们试图观察其对在含有干细胞因子（SCF）、白细胞介素3（IL-3）、白细胞介素6（IL-6）、粒细胞巨噬细胞集落刺激因子（GM-CSF）、促红细胞生成素（EPO）（Cys）以及MSP的组合或Cys与骨髓内皮细胞条件培养基（EC-CM）存在的情况下于液体培养系统中培养的早期造血祖细胞扩增的影响。
分离人骨髓CD34(+)细胞并在液体培养系统中培养6天。采用粒细胞巨噬细胞集落形成单位（CFU-GM）和粒细胞、红细胞、巨噬细胞、巨核细胞集落形成单位（CFU-GEMM）来检测不同处理对造血干/祖细胞增殖的影响。采用硝基蓝四氮唑（NBT）还原试验和Hoechst 33258染色分别反映细胞的分化和凋亡。
MSP在半固体培养中抑制CFU-GM和CFU-GEMM的增殖，且对CFU-GEMM的抑制作用强于CFU-GM。MSP抑制造血刺激因子诱导的早期造血祖细胞的分化。在液体培养系统中培养6天后，与0小时对照相比，Cys + EC-CM、Cys + MSP或Cys中的骨髓（BM）CFU-GEMM分别增加2.3倍或1.7倍或显著降低。
MSP作为一种造血抑制剂，抑制造血刺激因子诱导的早期造血祖细胞的分化，并使早期造血祖细胞在液体培养系统中扩增。
