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目的研究转HOXB4基因的人骨髓间充质干细胞(MSC)对脐血CD34+细胞体外扩增的支持作用。方法用病毒载体质粒转染包装细胞293T,收获病毒上清(VCM)感染MSC后,用潮霉素筛选获得转HOXB4的MSC(MSC-HOX)。分别将MSC(对照组)与MSC-HOX细胞(实验组)作为CD34+细胞体外扩增的饲养层细胞,结合细胞因子Flt3/Flk3配体(FL)、促血小板生成素(TPO)、干细胞因子(SCF)和粒细胞-集落生长因子(G-CSF)体外扩增脐血CD34+细胞10d,收集所有脐血细胞,检测细胞总数、CD34+细胞总数、集落细胞形成单位(CFU)数。结果生产重组逆转录病毒可有效将HOXB4基因转入靶细胞内并表达。脐血CD34+细胞经体外扩增10d后,细胞总数和CFU数两组间差异无统计学意义(P>0.05),脐血CD34+细胞总数和比例高于对照组(P<0.05)。结论转HOXB4基因的人骨髓MSC在脐血CD34+细胞体外扩增中,可保持CD34+细胞未分化状态,有潜在的应用价值。
Objective To investigate the effect of human bone marrow mesenchymal stem cells (HOXB4) transfected with HOXB4 gene on the expansion of cord blood CD34 + cells in vitro. Methods 293T cells were transfected with 293T virus vector and infected with virus supernatant (VCM). MSC infected with MSC were transduced with hygromycin (MSC-HOX). MSCs (control group) and MSC-HOX cells (experimental group) were used as feeder layer cells in vitro to proliferate CD34 + cells. FLT3 / Flk3 ligand (FL), thrombopoietin (TPO), stem cell factor SCF) and granulocyte-colony-forming growth factor (G-CSF) in vitro for 10 days. All the cord blood cells were collected. The total number of cells, the total number of CD34 + cells and the number of colony forming units (CFU) were detected. Results The production of recombinant retroviruses efficiently transfered the HOXB4 gene into target cells and expressed it. There was no significant difference between the two groups (P> 0.05). The total number and proportion of CD34 + cells in umbilical cord blood were higher than those in control group (P <0.05). Conclusion The human bone marrow MSCs transfected with HOXB4 gene can maintain the undifferentiated state of CD34 + cells in vitro in cord blood CD34 + cells, which has potential value.