建立一种以纤维蛋白凝胶为基底的体外血管模型

在组织工程构建体外血管样结构的基础上,建立一种以纤维蛋白凝胶为基底的体外血管再生模型并对其影响因素进行研究。采用人脐静脉内皮细胞,将其接种在纤维蛋白凝胶上,观察其形成血管血管样管腔结构。然后对纤维蛋白凝胶的条件(纤维蛋白原的浓度、凝血酶的浓度和细胞密度、血清浓度)进行优化。结果显示,当纤维蛋白原的浓度为0.5 mg/mL,凝血酶的浓度为0.1 U/mL,细胞密度为1.5×104个/孔,血清浓度为10%时,建立的血管模型形成的管腔结构最多最长。此体外血管模型的建立不需添加其它的血管细胞生长因子,方便、简单,可用于血管化的研究。

Fibrin-based model of the in vitro Angiogenesis for tissue engineering

The present pilot study aims to construct a possible optimal angiogenesis model in vitro on fibrin gel for tissue engineering. We mixed fibrinogen and thrombin to form fibrin gel that contained no stimulatory factors as a culture substratum, and then endothelial cells (HUVEC) as the model cell line were seeding on the fibrin matrix to develop a test model. The development of tube-like structure (TLS) was examined in-situ with a phase-contrast and the length of it was calculated by software. Results showed that tubular structure formed within 24 h on proper concentration of fibrinogen, which was also sensitive to the other surrounding environment (i.e. concentration of thrombin, blood serum and HUVEC). Furthermore, the TLS were formed more favorable for the concentration of fibrinogen (0.5mg/mL), thrombin (0.1 U/mL), blood serum (10%) and HUVECs (1.5mg/ml). The simplicity, reproducibility of the fibrin-based model described here is a useful in vitro angiogenesis model for the further study of tissue-engineering vascularization.