Objective To evaluate the feasibility of utilizing vascular cells combined with folded andframed culture model to develop completely autologous human tissue without using any scaffold material under the principles of ...Objective To evaluate the feasibility of utilizing vascular cells combined with folded andframed 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 ascending aorta (group A) and 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 Immol/L L-ascorbic acid 2-phos-phate. 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, 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 groups after additional four weeks. DNA assay indicated significantly lower 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 contained 82% and 42% that of human pericardium. Conclusion It is feasible to obtain completely autologous human cardiovascular tissue with the alternative new approach. Numerous issues including improvement of mechanical strength of neo-tissue remain to be investingated.展开更多
文摘Objective To evaluate the feasibility of utilizing vascular cells combined with folded andframed 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 ascending aorta (group A) and 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 Immol/L L-ascorbic acid 2-phos-phate. 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, 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 groups after additional four weeks. DNA assay indicated significantly lower 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 contained 82% and 42% that of human pericardium. Conclusion It is feasible to obtain completely autologous human cardiovascular tissue with the alternative new approach. Numerous issues including improvement of mechanical strength of neo-tissue remain to be investingated.
