CAJ | 학술논문

目的探索人脂肪干细胞(adipose-derived stem cells,ADSCs)对兔任意型皮瓣成活能力的影响.方法获得健康成人的脂肪,经体外分离、培养及传代至第5代,进行流式细胞术检测和三系分化能力的鉴定.于大白兔背部对称地设计2个蒂部在头侧的任意型皮瓣(长6 cm、宽2 cm),左、右侧分别为对照组和实验组.应用DiO荧光标记ADSCs,实验组(n=10)分5点注射2×106悬浮于0.5 ml无血清的DMEM培养基中的ADSCs,对照组(n=10)同样分5点注入无血清的DMEM0.5 ml,于术后7 d分别测量皮瓣成活面积;随机作冰冻切片,在激光扫描共聚焦显微镜下观察切片荧光显影情况;随机作常规切片HE染色,光镜下观察皮瓣组织毛细血管分布密度.统计学分析ADSCs在促进任意型皮瓣成活过程中的作用.结果与对照组相比,实验组皮瓣成活率明显提高(P<0.01),组织学检测证明实验组毛细血管增生明显增多.结论人ADSCs具有较好的免疫相容性,局部应用可明显提高皮瓣的成活率.
목적 탐색인지방간세포(adipose-derived stem cells,ADSCs)대토임의형피판성활능력적영향.방법 획득건강성인적지방,경체외분리、배양급전대지제5대,진행류식세포술검측화삼계분화능력적감정.우대백토배부대칭지설계2개체부재두측적임의형피판(장6 cm、관2 cm),좌、우측분별위대조조화실험조.응용DiO형광표기ADSCs,실험조(n=10)분5점주사2×106현부우0.5 ml무혈청적DMEM배양기중적ADSCs,대조조(n=10)동양분5점주입무혈청적DMEM0.5 ml,우술후7 d분별측량피판성활면적;수궤작빙동절편,재격광소묘공취초현미경하관찰절편형광현영정황;수궤작상규절편HE염색,광경하관찰피판조직모세혈관분포밀도.통계학분석ADSCs재촉진임의형피판성활과정중적작용.결과 여대조조상비,실험조피판성활솔명현제고(P<0.01),조직학검측증명실험조모세혈관증생명현증다.결론 인ADSCs구유교호적면역상용성,국부응용가명현제고피판적성활솔.
Objective To explore the effect of the adipose derived stem cells (ADSCs)on the survival of random pattern skin flap in rabbits. Methods ADSCs were isolated from fresh human fat and expanded in vitro for five passages. Then the characterization of ADSCs were determined by their CD marker profile and their ability to differentiate into osteogenic, adipogenic and chondrogenic lineages. On the back of the rabbits, two symmetric cephalic-based random pattern skin flaps were designed (6 cm × 2cm). The right flaps were used as experimental groups with the contra-side flaps as control group. Human ADSCs were pre-labeled before seeding with fluorescent 3, 30-dioctadecyloxacarbocyanine perchlorate (DiO) dye. The experimental flaps evenly received 5-point injection of 2 × 106Dio-labeled ADSCs resuspended in 0.5 ml of serum-free DMEM, while only 0. 5 ml medium was injected into the control flaps.Seven days later, the survival rate of flaps was evaluated. The flaps underwent frozen section and were observed under the laser scanning confocal microscope to detect the fluorescence imaging. Flaps also underwent HE staining and were observed under light microscope to detect the vascular density. Results Compared with control group, there was a significant increase of flap survival rate in the experimental group(P <0.01). Histological analysis also demonstrated a statistically significant increase in capillary density in the experimental group. Conclusions It suggests that ADSCs have a better immune compatibility and potential for enhancing the blood supply of random pattern skin flaps.