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本研究探讨基质细胞衍生因子(stromal cell derived factor-1,SDF-1)及其受体CXCR4在人脐血源基质细胞(human umbilical cord blood-derived stromal cells,hUCBSCs)促进巨核细胞增殖中的作用。以巨核细胞系HEL细胞作为研究对象,实验分HEL/hUCBSC共培养组、HEL/骨髓基质细胞(human bone marrow stromal cells,hBMSCs)共培养组和HEL悬浮培养组。应用ELISA法检测hUCBSC和hBMSC培养上清SDF-1水平,应用激光共聚焦和流式细胞仪检测HEL细胞CXCR4蛋白表达,RT-PCR检测CXCR4 mRNA表达。结果表明:hUCBSC高分泌表达SDF-1,其分泌高峰在培养第8天,稍迟于hBMSCs。与hUCBSCs共培养的HEL细胞,流式细胞仪和激光共聚焦检测均显示其CXCR4蛋白的表达较弱(p<0.05),且可在部分HEL细胞胞浆中发现红色点状荧光。RT-PCR检测结果显示,不同培养条件下HEL细胞CXCR4 mRNA表达无显著性差异(p>0.05)。结论:hUCBSC在分泌SDF-1和调控巨核细胞表达CXCR4方面起重要作用。
This study was designed to investigate the effects of stromal cell derived factor-1 (SDF-1) and its receptor CXCR4 on the proliferation of megakaryocytes by human umbilical cord blood-derived stromal cells (human umbilical cord blood-derived stromal cells) . The megakaryocytic HEL cells were used as the research object. The experiment was divided into HEL / hUCBSC co-culture group, HEL / MSCs hCMSCs co-culture group and HEL suspension culture group. The levels of SDF-1 in hUCBSC and hBMSC culture supernatant were detected by ELISA. The expression of CXCR4 protein in HEL cells was detected by laser scanning confocal microscope and flow cytometry. The expression of CXCR4 mRNA was detected by RT-PCR. The results showed that hUCBSC highly secretes SDF-1, and its secretion peak is on the 8th day of culture and slightly later than that of hBMSCs. The HEL cells co-cultured with hUCBSCs showed that the expression of CXCR4 protein was weak (p <0.05) by flow cytometry and confocal laser scanning microscope, and red dot fluorescence was observed in some HEL cells. The results of RT-PCR showed that there was no significant difference in CXCR4 mRNA expression in HEL cells under different culture conditions (p> 0.05). Conclusion: hUCBSC plays an important role in the secretion of SDF-1 and the regulation of CXCR4 expression in megakaryocytes.