外周血来源内皮祖细胞三维血管新生模型的建立及其特性分析

Establishment and Analysis of Three-Dimensional Angiogenesis Model of Endothelial Progenitor Cell from Peripheral Blood

目的观察内皮祖细胞(EPC)在血管新生中的作用及其生物学特性。方法取大鼠外周血分离出EPC,观察其在体外的培养和扩增情况,对培养的EPC进行检测,同时建立三维立体模型并分析。结果成功分离出外周血中的EPC,在平面培养时,各时间点经VEGF诱导的EPC(实验组)其增殖情况均优于无VEGF诱导的EPC(对照组),P<0.01。在由鼠尾胶原凝胶制成的三维基质中EPC向胶原基质内生长,1d内即可出现向胶原内的出芽及浸润,并逐渐形成分支样结构;实验组生长快,向胶原基质内浸润速度快、出芽快,管状结构粗大;而对照组生长慢、出芽慢,管状结构细小,向胶原内浸润的深度浅,网状结构稀疏,不完整;实验组各时相三维基质中新生血管数目均多于对照组(P<0.01)。结论鼠尾胶原凝液可以诱导EPC参与血管新生的迁移、增殖、发芽等步骤;EPC三维基质模型可用于新生血管的研究。VEGF能动员和诱导EPC促进血管新生。

Objective To establish the three diamension-model and to observe the contribution of endothelial progenitor cell （EPC） in the angiogenesis and its biological features. Methods EPC was obtained from the rats＇ peripheral blood. Its cultivation and amplification in vitro were observed, and the function of the cultural EPC in vitro was detected. The three diamension-model was established and analyzed. Results EPC was obtained from the peripheral blood successfully. The proliferation of the EPC which induced with VEGF （experimental group） was better than that without VEGF （control group） at every different phase （P〈0.01）. It was found that EPC grew into collagen-material from up and down in the three diamension-model, and its pullulation and infiltration into the collagen were seen on day 1 after cultivation. With the time flying, there were branch-like constructions which were vertical to the undersurface of collagen and interlaced to net each other. It showed that in experimental group the EPC grew fast, its infiltration and pullulation also were fast, the branch-like construction was thick. But in control group, the EPC grew slowly, infiltration and pullulation were slow, the branch-like construction was tiny and the depth of infiltration into collagen was superficial. The number of new vessels in experimental group was larger than that in the control group at every different phase （P〈0.01）. Conclusion Rat tail collagen can induce EPC involved in immigration, proliferation and pullulation in angiogenesis. The three-diamension model of EPC can be used to angiogenesis research. VEGF can mobilize and induce EPC to promote the angiogenesis.