摘要
目的:比较血管基质成分(st romal vascul ar fract i on,SVF)与体外扩增的脂肪来源干细胞(adi pose-deri ved st emcel l s,ASCs)对移植脂肪成活的促进作用。方法:用手术方法切取家兔腹股沟脂肪,进行ASCs体外分离培养;取第3代ASCs分别进行成脂、成骨诱导实验,CD29和CD31流式鉴定;制备SVF,进行CD29和CD31流式鉴定;脂肪移植裸鼠实验分为3组:SVF、ASCs、和空白对照组(DMEM/F12),每组4只裸鼠,沿背部脊柱两侧对称部位4个移植位点,每组共16个注射移植位点,每点0.3ml脂肪颗粒(adi pose granul e,AG)+0.2ml细胞成分;术后4m时取材称重、固定行HE染色观察移植脂肪组织结构,行CD31免疫组化染色观察新生血管及其密度。结果:家兔ASCs体外分离培养成功,为贴壁生长,第3代细胞形态均呈长梭形。成脂诱导实验油红O染色显示形成脂滴,成骨诱导实验茜素红染色显示形成钙化结节。流式鉴定显示SVF:CD29:17.0%,CD31:1.3%;ASCs:CD29:96.2%,CD31:3.8%。SVF、ASCs和空白对照组各组移植脂肪成活量分别为0.2096±0.0024g,0.1798±0.0033g,0.1350±0.0020g,两两比较差异均有统计学意义,P<0.05。HE染色显示SVF组移植脂肪成活较好,组织结构完整,脂滴大小均一,可见脂肪组织中间有丰富的血管存在;ASCs组移植脂肪成活尚可,组织结构尚完整,脂滴大小一般均一,可见脂肪组织中有结缔组织纤维间隔和新生血管形成;空白对照组结缔组织纤维间隔明显增多,脂滴大小不一,有少量较大空泡形成。各组移植脂肪新生血管密度分别32.6±2.1条/mm2,29.3±1.6条/mm2,23.3±1.9条/mm2,两两比较均有统计学意义,P<0.05。结论:新鲜的干细胞成分SVF比体外扩增的ASCs能更好的促进移植脂肪成活。
Objective To compare the effect of stromal vascular fraction （SVF） and adipose-derived stem cells （ASCs） on fat graft survival. Methods ASCs were derived from subcutaneous fat tissue of rabbit dorsa and isolated, cultured ex vivo. The third passage of ASCs were conducted on the induction to fat and bone, and flow cytometry of CD29 and CD31. After SVF had been prepared, it was performed flow cytometry of CD29 and CD31 too. There were 3 groups in nude mice experiment of fat transplantation： SVF, ASCs, and DMEM/F12 as a blank control. There were 16 injection points on the paravertebral dorsa of 4 nude mice in each group. The transplanted mixture consisted of 0.3ml adipose granule （AG） and 0.2ml cell components. 4 months after the transplantation, the mice were euthanized and the fat grafts were dissected out, weighed, and analyzed by HE staining and immunohistochemistry staining of CD31. Results ASCs were successfully isolated and cultured ex vivo, and the third passage of them were fibroblast-like adherent cells. ASCs can be induced to lipid droplets and mineralization nodes. Flow cytometry indicated that： SVF： CD29： 17.0%, CD31： 1.3%; ASCs： CD29： 96.2%, CD31： 3.8%. The survival of the fat grafts in each group of SVF, ASCs and blank control group were 0.2096±20.0024g, 0.1798±20.0033g, 0.1350±20.0020g, there existing statistical significance between multiple comparisons. HE staining showed that the fat graft in group of SVF exhibited good survival, integrity, uniform lipid droplet and rich blood vessels; fat graft in group of ASCs exhibited survival, less integrity, less uniform lipid droplets andconnective tissue septa; the blank control group exhibited some large bubbles, all sizes lipid droplets, and significant fibrous septa. In each group the vascular density was 32.6±22.1 capillaries/mm^2, 29.3±21.6 capillaries/mm2, 23.3±21.9 capillaries/mm^2, there existing statistical significance between multiple comparison. Conclusions Fresh stem cell components SVF are more helpful than ASCs expanded ex vivo for fat graft survival.
出处
《中国美容医学》
CAS
2013年第1期35-39,共5页
Chinese Journal of Aesthetic Medicine
基金
国家自然科学基金面上项目(30973132)
全军医学科研"十二五"项目(BWS11C061)
关键词
脂肪移植
基质血管成分
脂肪来源干细胞
血管新生
fat transplantation
stromal vascular fraction
adipose-derived stem cells
neovascularization
