应用生物反应器构建组织工程化血管的实验研究

目的探讨利用生物反应器体外构建具有完整结构的组织工程化血管的可行性。方法从6月龄毕格犬颈总动脉获取平滑肌细胞,颈外静脉获取内皮细胞,经体外培养扩增。然后,先将平滑肌细胞接种于聚羟基乙酸(PGA)上,形成细胞—材料复合物,将该复合物置于生物反应器内培养。模拟成年哺乳动物循环系统的参数,予以动态力学刺激(搏动频率:75次/分;扩张量<5%)。8周后,在其上接种内皮细胞,对照组不接种内皮细胞,培养5d后取材检测。结果组织学染色显示,平滑肌纤维成分排列较规则,有层次感,内皮细胞完整;对照组未见内皮细胞层结构。结论利用生物反应器可在体外构建具有完整结构的组织工程化血管。

Experimental Study on Tissue Engineered Blood Vessel with Bioreactor

Objective To investigate the feasibility of constructing tissue engineered blood vessels with bioreactor. Methods Cell-PGA （polyglycolic acid） complex was constructed by seeding smooth muscle cells （SMCs） isolated from canine carotid artery on PGA unwoven fibers. The cell-PGA complex was cultured in a bioreactor （ pulse rate： 75 /min, radical distension〈5%）. Eight weeks after culture in vitro, Endothelial cells （ECs） isolated from canine carotid vein were seeded on PGA and cultured for another 5 days. The control group was not seeded ECs. Tissue engineered blood vessels were harvested and examined. Results Histological staining revealed layered smooth muscle fibers and integral ECs in the tissue engineered blood vessel wall, which was further confirmed by immuno-histological staining. There were no obvious ECs were observed in the control group. Conclusion The technology of bioreactor could be used to build tissue engineered blood vessel with mimic native blood vessel.