Wendt D, Marsano A, Jakob M, Heberer M, Martin I
Departments of Surgery and of Research, University Hospital Basel, Hebelstrasse 20, ZLF, Room 405, 4031 Basel, Switzerland.
Biotechnol Bioeng. 2003 Oct 20;84(2):205-14. doi: 10.1002/bit.10759.
We developed a bioreactor for automated cell seeding of three-dimensional scaffolds by continuous perfusion of a cell suspension through the scaffold pores in oscillating directions. Using quantitative biochemical and image analysis techniques, we then evaluated the efficiency and uniformity of perfusion seeding of Polyactive foams as compared to conventional static and spinner flask methods. Finally, we assessed the efficacy of the perfusion seeding technique for different scaffolds and cell types. Perfusion seeding of chondrocytes into Polyactive foams resulted in "viable cell seeding efficiencies," defined as the percentages of initially loaded cells that were seeded and remained viable, that were significantly higher (75 +/- 6%) than those by static (57% +/- 5%) and spinner flask seeding (55% +/- 8%). In addition, as compared to static and spinner flask methods, cells seeded by perfusion were respectively 2.6-fold and 3.8-fold more uniformly distributed and formed more homogeneously sized cell clusters. Chondrocytes seeded by perfusion into Hyaff-11 nonwoven meshes were 26% and 63%, respectively, more uniformly distributed than following static and spinner flask seeding. Bone marrow stromal cells seeded by perfusion into ChronOS porous ceramics were homogeneously distributed throughout the scaffold volume, while following the static method, cells were found only near the top surface of the ceramic. In summary, we demonstrated that our cell seeding perfusion bioreactor generated constructs with remarkably uniform cell distributions at high efficiencies, and was effective for a variety of scaffolds and different mesenchymal cell types.
我们开发了一种生物反应器，通过在振荡方向上连续灌注细胞悬液，实现对三维支架进行自动细胞接种。然后，我们使用定量生化和图像分析技术，评估了与传统静态和旋转瓶方法相比，聚活性泡沫灌注接种的效率和均匀性。最后，我们评估了灌注接种技术对不同支架和细胞类型的效果。将软骨细胞灌注接种到聚活性泡沫中，产生了“活细胞接种效率”，定义为接种并保持存活的初始加载细胞的百分比，该效率显著高于静态接种（57%±5%）和旋转瓶接种（55%±8%），达到75%±6%。此外，与静态和旋转瓶方法相比，灌注接种的细胞分布均匀度分别提高了2.6倍和3.8倍，并且形成了大小更均匀的细胞簇。将软骨细胞灌注接种到Hyaff-11无纺布网片中，其分布均匀度分别比静态和旋转瓶接种提高了26%和63%。将骨髓基质细胞灌注接种到ChronOS多孔陶瓷中，细胞均匀分布在整个支架体积内，而采用静态方法时，细胞仅出现在陶瓷的顶面附近。总之，我们证明了我们的细胞接种灌注生物反应器能够高效地产生细胞分布非常均匀的构建体，并且对多种支架和不同间充质细胞类型均有效。
