Oscillating perfusion of cell suspensions through three‐dimensional scaffolds enhances cell seeding efficiency and uniformity

TLDR

The study aimed to develop an automated bioreactor that perfuses a cell suspension through scaffold pores in oscillating directions and to evaluate its efficacy across multiple scaffold types and mesenchymal cell lines. The bioreactor delivers continuous oscillatory perfusion of cells into three‑dimensional scaffolds, and the authors assessed seeding efficiency and uniformity using quantitative biochemical assays and image‑based analysis. Perfusion seeding achieved 75 % viable cell efficiency in Polyactive foams, 2.6‑ to 3.8‑fold greater uniformity than static or spinner methods, and produced homogeneous cell distributions in Hyaff‑11 and ChronOS scaffolds, outperforming static seeding which yielded uneven, surface‑localized cell placement. © 2003 Wiley Periodicals, Inc., Biotechnol Bioeng 84:205–214.

Abstract

Abstract 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. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 205–214, 2003.