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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... 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology and Bioengineering Wiley

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

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References (41)

Publisher
Wiley
Copyright
Copyright © 2003 Wiley Periodicals, Inc., A Wiley Company
ISSN
0006-3592
eISSN
1097-0290
DOI
10.1002/bit.10759
pmid
12966577
Publisher site
See Article on Publisher Site

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.

Journal

Biotechnology and Bioengineering – Wiley

Published: Oct 20, 2003

Keywords: cell seeding; bioreactors; tissue engineering; three‐dimensional scaffolds; image analysis

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