Method to Analyze Three-Dimensional Cell Distribution and Infiltration in Degradable Scaffolds
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Date
2008Author
Thevenot, Paul Todd
Nair, Ashwin
Dey, Jagannath
Yang, Jian
Tang, Liping
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**Please note that the full text is embargoed** ABSTRACT: Effective cell seeding throughout the tissue scaffold often determines the success of tissue-engineering products,
although most current methods focus on determining the total number, not the distribution, of the cells associated
with tissue-engineering constructs. The purpose of this investigation was to establish a quick, convenient, and
efficient method to quantify cell survival, distribution, and infiltration into degradable scaffolds using a combination of fluorescence cell staining and cryosectioning techniques. After cell seeding and culture for different
periods of time, seeded scaffolds were stained with a live cell dye and then cryosectioned. Cryosectioned scaffolds
were then recompiled into a three-dimensional (3D) image to visualize cell behavior after seeding. To test the
effectiveness of this imaging method, four common seeding methods, including static surface seeding, cell injection, orbital shaker seeding, and centrifuge seeding, were investigated for their seeding efficacy. Using this new
method, we were able to visualize the benefits and drawbacks of each seeding method with regard to the cell
behavior in 3D within the scaffolds. This method is likely to provide useful information to assist the development
of novel materials or cell-seeding methods for producing full-thickness tissue grafts.