人脐血来源内皮祖细胞促进裸鼠皮瓣存活的实验研究

The study on EPC originated from human umbilical cord blood promoting neovascularization in ischemic skin flap

目的将内皮祖细胞（endothelial progenitor cell，EPC）引入皮瓣缺血组织中，初步研究其生物学特性及在皮瓣缺血组织中促血管发生的作用。方法采用免疫磁珠法从人脐血中分离培养CD133＋内皮祖细胞，将体外扩增的EPC局部注射于裸鼠超长皮瓣模型中，观察EPC转归及参与皮瓣血管重建的情况；通过观察皮瓣坏死面积及微血管增生情况，评价EPC在皮瓣缺血组织中再血管化能力。结果EPC组皮瓣坏死面积明显小于空白组（P〈0．05）、真皮下层组织灌流量大于对照组（P〈0．05）；荧光示踪及免疫组织化学染色证实EPC参与皮瓣血管重建。结论EPC参与血管重建，具有促进血管新生，加速缺血组织血管化的作用。

Objective Injecting the EPC into the corresponding skin flap to study EPC biological characteristics and its effect on neovascularization in ischemia skin flap. Methods CD133 ＋ cells were enriched from human umbilical cord blood by immunomagnetic sorting, and cultured with EGM - 2MV media. After labeled with PKH26 （fluorescent cell linker）, the EPC were injected into the over-length flap models made on athymie mice. Observing the EPCs trace and their participating in the flap vascularization using a fluorescent microscope. The potential of EPC neovascularization in ischemic tissue of skin flap was evaluated through measuring the necrotic area and vessel diameter and quantity in the skin flap. Results The skin flap necrosis area of EPC group is significantly smaller than that of control （ P 〈 0.05 ）, the dermal and hypodermal blood perfusion of EPC group is significantly more than that of control （ P 〈 0.05 ）. Immunohistological and label fluorescent analyses showed vWF antigen-positive cells and labeled cells constructing blood vessels of flap. Conclusions Our data support the EPC may contribute to angiogenesis, speed up ischemic tissue vascularization.