内皮细胞流动培养模型的建立

Establish Cultured Model of Endothelial Cells in a Flow Environment

目的 促进培养内皮细胞 (EC)分化 ,增强 EC对流体剪切力的抵抗。 方法 利用改进的流室装置 ,通过精密蠕动泵提供剪切力 ,同时提供静态培养所需的其它条件 ,建立体外的 EC流动培养模型 ,按流动培养增加剪切力的方式分为 A组、B组和 C组 ,并筛选出最佳的流动培养模式。 结果 EC在流动培养下生长良好 ,血管 EC被拉长 ,其长轴方向与流场方向趋于一致。不同流动培养方式作用下 ,EC能适应剪切力递增式 (A组 )和阶跃式 (B组 )增加的作用 ,直接暴露于较大的剪切力作用下发生较明显的细胞脱落 (C组 )。 结论 我们建立的模型能相对地接近模拟体内环境 ,阶跃式增加剪切力在流动培养中方便、切实可行。

Objective\ To promote the differentiation of cultured endothelial cells and enhance their resistance to fluid shear stress.\ Methods\ Using the mended parallel plate flow apparatus and peristalsis pump providing fluid shear stress, endothelial culture models were established in vitro with the same environment factors as steady culture. According to the increasing degree of shear stress, the experiment included:(1) Group A, exposing to the gradual increasing fluid shear stress, (2) Group B, exposing to step wise fluid shear stress, and (3) Group C, directly exposing to the fluid shear stress 1.5 Pa for 48 hours. The best culture model was selected.\ Results\ The cultured endothelial cells under flow condition grew well. The cells elongated with the longitudinal axis paralleled the direction of flow. Under different culture mode, the endothelial cell adapted to gradual increasing or step wise increasing fluid shear stress(group A and B)but detached from the graft exposing to acute fluid shear stress(group C). Conclusion\ The cultured mode established can relatively mimic the environment in vivo, step wise increasing fluid shear stress is convenient and feasible for endothelial cell culture.