Centrifugal seeding of mammalian cells in nonwoven fibrous matrices.
Three-dimensional (3D) cell cultures have many advantages over two-dimensional cultures. However, seeding cells in 3D scaffolds such as nonwoven fibrous polyethylene terephthalate (PET) matrices has been a challenge task in tissue engineering and cell culture bioprocessing. In this study, a centrifugal seeding method was investigated to improve the cell seeding efficiency in PET matrices with two different porosities (93% and 88%). Both the centrifugal force and centrifugation time were found to affect the seeding efficiency. With an appropriate centrifugation speed, a high 80-90% cell seeding efficiency was achieved and the time to reach this high seeding efficiency was less than 5 min. The seeding efficiency was similar for matrices with different porosities, although the optimal seeding time was significantly shorter for the low-porosity scaffold. Post seeding cell viability was demonstrated by culturing colon cancer cells seeded in PET matrices for over 5 days. The centrifugal seeding method developed in this work can be used to efficiently and uniformly seed small fibrous scaffolds for applications in 3D cell-based assays for high-throughput screening.
William G. Lowrie Dept. of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA.
Robin
R
Ng
Jesse Singh
JS
Gurm
Shang-Tian
ST
Yang
D018929:Cell Culture Techniques / Q000379:methods*; D002498:Centrifugation; D005109:Extracellular Matrix / Q000737:chemistry*; D057166:High-Throughput Screening Assays; D006801:Humans; D011093:Polyethylene Terephthalates / Q000737:chemistry*; D016062:Porosity; D049268:Positron-Emission Tomography; D013499:Surface Properties; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D014407:Tumor Cells, Cultured
D011093:Polyethylene Terephthalates