接种方式对细胞在三维材料中接种率和分布的影响

考察了静态和动态接种方式对成纤维细胞在胶原壳聚糖支架材料中接种率和分布的影响。将人成纤维细胞制成细胞悬液，分别采用静态接种、转瓶接种和灌注接种方式将细胞接入三维胶原壳聚糖海绵。通过MTT法和切片HE染色分别考察细胞接种率及细胞在三维材料中的分布。实验结果表明：在低的接种密度下静态接种有较高的接种率(889%)，但随着接种密度的增加接种率下降显著，细胞结团且分布不均匀；转瓶接种的接种率约为60%，细胞分布也不均匀；灌注接种的接种率始终维持在77%以上，能得到高的起始细胞密度，且细胞分布均匀，是一种理想的接种方式。细胞接种方式的优化为改善工程化组织的结构和功能、缩短体外构建时间奠定了基础。

Effects of Seeding Methods on Seeding Efficiency and Initial Cell Distribution in 3-D Scaffolds

Cell seeding of three-dimensional scaffolds is the first step of the cultivation of engineered tissues. The cell seeding density and spatial distribution in a 3-D scaffold are critical to the morphogenetic development of an engineered tissue. In the present work, human fibroblasts were seeded to collagen-chitosan sponges by static seeding, stirred seeding and perfusion seeding. The effects of seeding conditions on the resulting seeding density, the seeding efficiency and the initial cell distribution were studied. The seeding efficiency was relatively high(88.9%) at low inoculation cell density, and decreased rapidly with the increasing of inoculation cell density in static seeding. Stirred seeding yielded the lowest seeding efficiency. Nonuniform initial cell distribution was observed in both static and stirred seeding. The perfusion seeding, which has a characteristic of high seeding efficiency(>77%) , high initial cell density and uniform initial cell distribution in 3-D scaffolds, is the optimum method for cell seeding to 3-D scaffold.