培养方式对真皮组织体外构建的影响

采用静态培养和转瓶培养方式分别构建真皮组织，考察培养方式和搅拌转速对细胞在三维支架材料中增殖、代谢、分布的影响。结果表明，由转瓶培养方式构建的细胞-材料复合物，其最终细胞密度和细胞比生长速率均明显高于静态培养（14.2~27.6×106 cells/cm3 vs 10.1×106 cells/cm3和0.145~0.262 d-1 vs 0.111 d-1），而转速达80 r/min的转瓶尤其突出；静态培养的细胞-材料复合物内部细胞稀少，且分布不均匀，转瓶培养的细胞-材料复合物在材料表面和内部细胞密度都有所提高，分布情况也得到改善，且80 r/min转瓶培养的组织其细胞密度和分布均优于10 r/min和40 r/min转瓶培养。转瓶培养在其转速达到一定强度时能明显提高细胞在支架中的增殖速率，缩短培养时间，并有效改善细胞在支架内的分布，是一种理想的培养方式。

Effects of the culture method on the construction of dermal substitutes in vitro

Culture environment is the key factor in the construction of dermal skin. This study reports on the effects of culture methods on the construction of dermal substitutes using collagen-chitosan sponges in vitro. Through comparing the static culture and spinner flask culture methods, and the spinner flask culture with different stir speeds 10 r/min, 40 r/min and 80 r/min, the effects of the culture environment on the cells proliferation, cells distribution within scaffold and cells metabolism had been studied. A greater cell density was obtained with spinner flask culture versus static culture, especially, the 80 r/min spinner flask culture shows the obvious advantage with the greatest cell density and specific growth rate. The stirred culture of fibroblast-seeded sponges in spinner flasks resulted in a more uniform cell distribution compared to static culture. The dermal substitutes obtained from the 80 r/min spinner flask culture have greater cell density and more uniform distribution within scaffold than the 10 r/min and 40 r/min spinner flaks. In summary, the spinner flasks culture with proper stir speed shows promise for the in vitro construction of dermal substitutes.