体外扩增恒河猴自体内皮细胞人工血管移植研究

目的为自体内皮细胞移植的临床应用奠定理论基础。方法将恒河猴表浅静脉内皮细胞在体外培养１３．９±１．４天;扩增培养的自体细胞衬里于用纤维蛋白胶预衬的膨体聚四氟乙烯（ｅＰＴＦＥ）人工血管,继续培养９天。１５只恒河猴随机分为两组。实验组（１０只）用衬有自体静脉内皮细胞的人工血管置换骼总动脉;对照组（５只）用未衬里内皮细胞的人工血管置换。结果细胞数增加１４８±８９倍。所培养的细胞为二倍体细胞,纯度９９％。原代及传代细胞培养上清液中６－酮－前列腺素Ｆ１ａ（６－ｋｅｔｏ－ＰＧＦ１ａ）和ＶｏｎＷｉｌｅｂｒａｎｄ因子（ｖＷＦ）含量差异无显著意义。细胞种植后２小时和９天,人工血管腔面见一层均匀的基质,基质表面有一层连续的内皮细胞单层。内皮细胞紧密排列。呈梭状,形态饱满。实验组除１只于术后１２小时因近端吻合口破裂腹腔内出血死亡,１只人工血管于术后４周阻塞外,其余８只恒河猴人工血管术后４周均保持通畅,内膜下组织厚度为８０±１２μｍ;对照组５条人工血管全部阻塞。结论自体内皮细胞移植可有效地提高人工血管的通畅率。

Autologous endothelial cell transplantation in meacacos

OBJECTIVE: To lay a theoretical foundation for the clinical application of autologous endothelial cell transplantation. METHODS: Macaco endothelial cells derived from superficial veins were cultivated in vitro for 13.89 +/- 1.36 days. The multiplied cells were lined in vitro onto the luminal surface of expanded polytetrafluoroethylene (ePTFE) grafts precoated with fibrin glue and fibronectin, then cultivated further for 9 days. 15 macacos were divided randomly into two groups. The common iliac arteries of the exper mental animals (n = 10) were replaced with endothelialized grafts, those of control (n = 5) with untreated ones. RESULTS: The amount of endothelial cells increased for 147.93 +/- 88.68 folds. All the cells were diploid cells with a purity of 99%. The content of both 6-keto-PGF1a and vWF in the supernatant of primary and subcultured passages didn't have significant difference. Two hours and 9 days after cell seeding, the luminal surface of grafts were covered completely by a prominent, spindle-like endothelial monolayer underneath an even fibrin glue matrix could be seen. Nine days after seeding, the dense condensation of cytoskeleton on the luminal and basal side of cells increased apparently. Four weeks after grafts implantation 8/10 of experimental grafts, with a thickness of intima 80 +/- 12 microns, were patent; while the 5 control grafts were occluded. CONCLUSION: The endothelial cell transplantation could effectively increase the patency rate of synthetic blood vessel prosthesis.