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采用密度梯度离心和差速贴壁法分离培养人骨髓基质细胞。对细胞进行形态观察、表型分析、细胞生长曲线和成骨分化潜能测定。然后将第4代细胞通过尾静脉输注到未经任何处理的裸鼠体内,移植1周和4周后,利用RT-PCR检测人Alu基因在裸鼠不同脏器组织中的表达以判断人源性供体细胞的植入情况。结果表明培养的人骨髓基质细胞为CD34-CD45-成纤维样细胞,并具有向成骨分化的能力,但第1至第5代人骨髓基质细胞增殖速度无显著性差异(P>0.05)。经尾静脉输注后,人源性供体细胞可在肺、肝、脾、肾、肌肉、心、肠、脑等多种脏器或组织中均有不同程度的分布。
Human bone marrow stromal cells were isolated and cultured by density gradient centrifugation and differential adherence. The cells were observed morphologically, phenotype analysis, cell growth curve and osteogenic differentiation potential determination. The fourth generation cells were then transfused into the untreated nude mice via the tail vein. After 1 week and 4 weeks of transplantation, the expression of human Alu gene in different organs of nude mice was detected by RT-PCR to judge the human Implantation of derived donor cells. The results showed that the cultured human bone marrow stromal cells were CD34-CD45-fibroblast-like cells and had the ability to differentiate into osteoblasts. However, there was no significant difference in the proliferation rate of the first to fifth generation human bone marrow stromal cells (P> 0.05). After the tail vein infusion, human-derived donor cells can have different degrees of distribution in many organs or tissues such as lung, liver, spleen, kidney, muscle, heart, intestine and brain.