Oscillating perfusion of cell suspensions through three-dimensional scaffolds enhances cell seeding efficiency and uniformity
Oscillating perfusion of cell suspensions through three-dimensional scaffolds enhances cell seeding efficiency and uniformity
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DOI：
10.1002/bit.10759
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发表时间：
2003-10-20
影响因子：
3.8
通讯作者：
Martin, I
中科院分区：
文献类型：
--
作者：
Wendt, D;Marsano, A;Martin, I
我们开发了一种生物反应器，通过在振荡方向上通过支架孔连续灌注细胞悬浮液来自动接种三维支架。使用定量生物化学和图像分析技术，我们然后评估的效率和均匀性灌注接种的聚活性泡沫相比，传统的静态和旋转瓶的方法。最后，我们评估了灌注接种技术对不同支架和细胞类型的有效性。将软骨细胞灌注接种到Polyactive泡沫中导致“活细胞接种效率”，定义为接种并保持活力的初始加载细胞的百分比，其显著高于静态（57% +/- 5%）和转瓶接种（55% +/- 8%）。此外，与静态和转瓶方法相比，通过灌注接种的细胞分别是2.6倍和3.8倍更均匀地分布，并形成更均匀大小的细胞簇。通过灌注接种到Hyaff（R）-11非织造网中的软骨细胞分别为26%和63%，比静态和转瓶接种后的分布更均匀。通过灌注接种到ChronOS（TM）多孔陶瓷中的骨髓基质细胞均匀地分布在整个支架体积中，而按照静态方法，仅在陶瓷的顶面附近发现细胞。总之，我们证明了我们的细胞接种灌注生物反应器以高效率产生具有显著均匀的细胞分布的构建体，并且对各种支架和不同的间充质细胞类型有效。(C)2003 Wiley Periodicals，Inc.
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(R)-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(TM) 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. (C) 2003 Wiley Periodicals, Inc.