新鲜分离的脂肪SVF细胞促进脂肪移植存活的实验研究

目的 探讨应用自脂肪组织新鲜分离的血管基质层细胞辅助脂肪移植,提高移植物存活率的可行性.方法 将0.5 ml待移植的脂肪颗粒分别与下列细胞混合:①DiI标记的新鲜分离的自体血管基质层细胞(A组);②DiI标记的培养至第4代自体脂肪来源干细胞(B组);③DMEM完全培养基(C组),随机注射移植于14只新西兰兔背部皮下.术后观察:①湿重;②切片HE染色计数血管密度;③方网测试系统"点计数"法检测存活脂肪细胞计数以及纤维组织计数;④荧光显微镜检测DiI标记的细胞在体内的分化转归.结果 ①湿重:A组(291.0±72.1)mg,B组(269.3±67.3)mg,C组(177.8±60.0)mg,A、B两组脂肪存活率均高于C组(P<0.05),两组之间比较差异无统计学意义(P>0.05);②血管密度:A、B两组血管密度均高于C组(P<0.05),两组之间比较差异无统计学意义(P>0.05).③点计数:A、B两组存活脂肪细胞计数均高于C组(P<0.05),纤维组织计数均低于C组(P<0.05),两组之间比较差异均无统计学意义(P>0.05);④荧光显微镜下观察发现自体血管基质层细胞与自体脂肪来源干细胞在体内均可向血管内皮细胞分化.结论 自体血管基质层细胞与培养的自体脂肪来源于细胞均可提高脂肪移植物存活率,但前者操作更方便,安全性更高,具有广阔的临床应用前景.

Experimental study of the effect of adipose stromal vascular fraction cells on the survival rate of fat transplantation

Objective To investigate the effect of adipose stromal vascular fraction cells (SVFs)on the survival rate of fat ransplantation. Methods 0. 5 ml autologous fat tissue was mixed with: ①DiI-labeled autologous SVFs ( Group A ) ; ② DiI-labeled autologous adipose-derived stem cells ( ASCs ) ( Group B) ; ③Complete DMEM( Group C). And then the mixture was injected randomly under the back skin of 14 rabbits. The transplanted fat tissue in three groups was harvested at 6 months after implantation. Wet weight of fat grafts was measured for macroscopic aspects. After HE staining, blood vessel density, viable adipocytes and fibrous proliferation were counted respectively for histological evaluation. Trace of DiI-labeled ASCs in vivo was detected by fluorescent microscope. Results The wet weight of fat grafts in group A ( 291.0±72.1 )mg and group B ( 269.3±67.3 )mg was significantly higher than that in group C (177.8±60.0) mg, but the differnce between Group A and Group B was not significant. Histological analysis revealed that the fat grafts in group A and B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis, compared with the fat grafts in group C. The grafts in both group A and B had significantly higher capillary density than those in the control group. Part of vascular endothelial cells were observed to origin from ectogenic DiI-labeled SVFs and ASCs. Conclusions The autologous isolated SVFs has a similar effect as autologous cultured ASCs to improve the survival rate of fat transplantation. And the former is more practical and safe, indicating a wide clinical application in the future.