体外培养心血管组织的研究

The Study of Cardiovascular Tissue Culture in vitro

目的探讨以血管细胞结合折叠-支架培养模式来获得完全自身心血管新生组织的可行性。 方法将人体大隐静脉和升主动脉血管壁组织培养出的细胞种植于15cm培养碟中,经过4周的培养,将获得的细胞薄膜折叠成4层,固定于支架上,再经过4周的培养,获得来源于静脉或动脉血管壁的完全自身组织。比较二种来源的新生组织的组织学、超微形态学、DNA含量、胶原含量的差异。 结果经过4周的培养,二种来源的细胞都形成含有多层细胞的薄膜;再4周的培养后,折叠-支架培养模式得到的组织比非折叠-支架培养的组织更有弹性,形态学上更均一致密,有更多的细胞外基质生成,虽然DNA含量低于后者,但胶原含量明显高于后者;动脉和静脉来源的细胞经折叠-支架培养得到的组织,其胶原含量分别达到人体心包的82％和42％。 结论仅以血管细胞结合折叠-支架培养模式来获得完全自身心血管新生组织的方法是可行的。

Objective To evaluate the feasibility of utilizing vascular cells combined with folded and framed culture model to develop completely autologous human tissue without using any scaffold material under the principles of tissue engineering. Methods Human vascular cells cultured from the ascending aorta (group A) and the saphenous vein (group B) were seeded into 15cm -dishes (each n = 12) and cultured to form cell sheets over a period of four weeks with Dulbecco' s modified Eagle' s medium supplemented with 1 mmol/L L -ascorbic acid 2 -phosphate. Thereafter, cell sheets (6 samples of each group) were four - layer folded and cultured in a newly developed frame device for additional four weeks. Controls remained under standard culture conditions. Tissue development was evaluated by light and electron microscopy and biochemical assays. Results The formation of multi - layered cell sheets and production of extracellular matrix were observed in each group after the initial four weeks. Analysis of the folded and framed neo - tissue revealed a solid structure with increased matrix formation and tissue organization compared to the control group after additional four weeks. DNA assay indicated significantly less cell proliferation in folded and framed cell sheets than in that of unframed counterparts. Yet hydroxyproline assay demonstrated significant increase of collagen content in the framed aortic and venous derived tissues, which were 82% and 42% of the human pericardium. Conclusion It was feasible to obtain completely autologous human cardiovascular tissue with the new alternative approach.