目的:探讨经耳源静脉移植入体内的骨髓间充质干细胞(Bone marrow-derived mesenchymal stem cells,BM-MSCs)参与皮肤扩张的情况.方法:以小型家猪为研究对象,分离培养并鉴定猪BM-MSCs.制作猪皮肤扩张模型,8头猪随机分为两组,每组4头,实验组(A组):扩张后耳源静脉注入DiI标记的猪BM-MSCs;对照组(B组):扩张后耳源静脉注入等量PBS,分别于扩张后7、14、28天测量两组扩张面积,冰冻切片观察标记细胞在扩张皮肤中的比例和分布情况,Real-time PCR检测基质细胞衍生因子-1(SDF-1)表达情况.结果:流式细胞仪鉴定细胞CD29、CD90表达阳性,CD34、CD45表达阴性,细胞标记率＞99%.扩张后第14天、28天实验组扩张面积较对照组明显增加(P＜0.05),冰冻切片显示,14天时实验组扩张皮肤可见DiI标记细胞聚集.PCR示SDF-1表达量实验组是对照组的5.8倍.结论:经静脉移植的BM-MSCs可迁移至扩张局部,并能有效促进皮肤扩张.%Objective To investigate the feasibility of transplantation of mesenchymal stem cell from ear vein into the skin and soft tissue expansion model. Methods MSCs were isolated from porket's bone marrow and cultured in vitro. Eight pigs were randomly divided into two groups: The group A was injected with MSCs which dyed with CM-Dil from ear vein; the group B was injected with PBS from ear vein as the control group. The expansion area was measured on day7,14,28 after operation and the proportion and distribution of labeled cells were detected by frozen section . Realtime quantitative PCR was used to quantify the expression of chemotactic growth factor- 1(SDF-1). Results The MSCs cultured expressed CD90 and CD29 highly but didn't express CD34 or CD45. The mark rate>99%. On day 14 and 28, the area of experimental group was increased significantly.compared to the control group (P<0.05). The result of frozen section show that the Dil-labled cells were gathered together in the experimental group on day 14 after expansion. Real-time quantitative PCR show that the target gene expression of SDF-1 was 5.8-fold higher than the control group. Conclusion BM-MSCs transplantated from ear vein can migrate to the local of expansion and promote the skin expansion.
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