骨髓细胞不同组份形成成纤维细胞集落形成单位的效率及1,25(OH)2D3和PGE2在其中的调节作用

Efficiency of colony forming unit-fibroblastic formed by the different fraction of bone marrow cells and regulation of 1,25(OH)_2D_3 and PGE_2 in colony formation

目的:研究骨髓间充质干细胞(BM-MSCs)是否存在于骨髓不同组份中,以及其成纤维细胞集落形成单位(CFU-f)形成效率是否受1,25二羟基维生素D3[1,25(OH)2D3]和前列腺素E2(PGE2)的调节。方法:将总骨髓细胞和培养1天后的非黏附骨髓细胞经梯度密度离心分为单核细胞和粒细胞/红细胞两组份或单核细胞、粒细胞和红细胞三组份,在缺乏或添加10-8 mol/L 1,25(OH)2D3或10-7 mol/L PGE2条件下贴壁培养10天,行亚甲基蓝染色,计数CFU-f数目。结果:①单核细胞组份CFU-f形成效率约占总CFU-f的10%,粒细胞/红细胞组份、粒细胞组份、红细胞组份CFU-f形成效率分别约占总CFU-f的90%、47%、35%,均明显高于单核细胞组份;②非黏附骨髓细胞来源的不同组份CFU-f形成效率占总CFU-f的71%,较总骨髓来源不同组份CFU-f形成效率明显增加;③1,25(OH)2D3和PGE2能够明显刺激总骨髓来源的单核细胞和粒细胞组份的CFU-f形成效率以及非黏附骨髓细胞来源的单核细胞、粒细胞和红细胞组份的CFU-f形成效率。结论:骨髓细胞各组份中都存在BM-MSCs,1,25(OH)2D3和PGE2都具有促进BM-MSCs扩增的作用。

Objective： To determine whether bone marrow mesenchymal stem cells（BM-MSCs） are presented in the different frac-tions of bone marrow cells and whether the efficiency of colony forming unit-fibroblastic（CFU-f） formed by BM-MSCs is regulated by 1,25（OH）2D3 and prostaglandin E2（PGE2）.Methods：Total bone marrow cells or non-adherent bone marrow cells derived from total bone marrow cell cultures for 1 day were separated into the mononuclear cell fraction and the granulocyte / erythrocyte fraction or the mononuclear cell fraction,the granulocyte fraction and the erythrocyte fraction by density-gradient centrifugation and were cultured in the absence or presence of 10-8mol/L 1,25（OH）2D3 or 10-7 mol/L PGE2 for 10 days.The resulting cells were stained with methylene blue and the number of CFU-f was counted.Results： ①The CFU-f formed by mononuclear cell fraction accounted for about 10% of the total CFU-f,and CFU-f formed by granulocyte / erythrocyte fraction,granulocyte fraction and erythrocyte fraction accounted for about 90%,47% and 35% of the total CFU-f,respectively;②The fractions derived from the non-adherent bone marrow cells ac-counted for about 71% of the total CFU-f,which was more than those formed by the fractions derived from total bone marrow cells;③ Treatment of 1,25（OH）2D3and PGE2 enhanced the CFU-f formation of the mononuclear fraction and the granulocyte fraction de-rived from total bone marrow cells and the mononuclear cell fraction,the granulocyte fraction and the erythrocyte fraction derived from non-adherent bone marrow cells.Conclusion： Our results indicate that BM-MSCs are presented in the different fractions of bone marrow cells,and 1,25（OH）2D3 and PGE2 play a role in stimulating proliferation of BM-MSCs.