Yang T H, Miyoshi H, Ohshima N
Department of Biomedical Engineering, Institute of Basic Medical Sciences, University of Tsukuba, Ibaraki 305-8575, Japan.
J Biomed Mater Res. 2001 Jun 5;55(3):379-86. doi: 10.1002/1097-4636(20010605)55:3<379::aid-jbm1026>3.0.co;2-z.
Cell seeding is one of the key procedures in the construction of tissue-engineered organs. In our previous efforts to create a bioartificial liver, high-density cultures of hepatocytes (>1 x 10(7) cells/1 cm(3)-substrate) and long-term maintenance of metabolic function were achieved with a packed-bed reactor utilizing porous poly(vinyl formal) (PVF) resin as a scaffold. However, a low seeding efficiency of about 30% remains a major obstacle to the scaleup of the reactor. In the present study, a new cell seeding method, centrifugal cell immobilization (CCI), which is based on alternating centrifugation and resuspension, was used to achieve high-density seeding and improve the seeding efficiency. Using the CCI method, the maximum density of the immobilized hepatocytes reached 3.8 x 10(7) cells/1 cm(3)-PVF, and the seeding efficiency was improved to about 43% after a relatively short immobilization process (about 15 min). Moreover, further improvement of the seeding efficiency was obtained by serial immobilization procedures. Thus, we concluded that this method is useful and effective for seeding cells into 3-dimensional scaffolds.
细胞接种是组织工程器官构建中的关键步骤之一。在我们之前构建生物人工肝的工作中，利用多孔聚（乙烯醇缩甲醛）（PVF）树脂作为支架的填充床反应器实现了肝细胞的高密度培养（>1×10⁷细胞/1 cm³底物）以及代谢功能的长期维持。然而，约30%的低接种效率仍然是该反应器扩大规模的主要障碍。在本研究中，一种基于交替离心和重悬的新细胞接种方法——离心细胞固定化（CCI）被用于实现高密度接种并提高接种效率。使用CCI方法，固定化肝细胞的最大密度达到3.8×10⁷细胞/1 cm³ - PVF，并且在相对较短的固定过程（约15分钟）后接种效率提高到约43%。此外，通过连续固定化程序进一步提高了接种效率。因此，我们得出结论，该方法对于将细胞接种到三维支架中是有用且有效的。
