Measuring Cell Adhesion and Proliferation in Polymer Scaffolds by X-Ray Microcomputed Tomography
Shauna M. Dorsey, Sheng Lin-Gibson, Carl Simon Jr.
We have explored a new X-ray microcomputed tomography method (CT) for assessing cell adhesion and proliferation in polymer scaffolds. Standard methods for examining cells in scaffolds include fluorescence microscopy and soluble assays for cell components such as enzymes, protein or DNA. Fluorescence microscopy is generally qualitative and cannot visualize the scaffold interior. Soluble assays quantitatively measure cell number but do not yield images. In contrast, CT enables quantitative, three-dimensional (3D) imaging of cells throughout the scaffold. Herein, the ability of CT to detect cells in scaffolds was compared with fluorescence microscopy and a soluble assay for DNA (Picogreen). Cells were seeded into polymer scaffolds and the measurement of their adhesion and proliferation using three approaches, fluorescence microscopy, Picogreen DNA assay and CT, was compared. The results showed that greater than 25000 cells had to be seeded on a 96-well scaffold (6.5 mm dia.) in order to enable detection by CT. Fluorescence microscopy had better resolution than mCT, but can not image cells through the scaffold struts. The Picogreen assay was more sensitive than CT but did not yield images. These results demonstrate that mCT can be used to detect cells in polymeric scaffolds for quantitative 3D evaluation of cell adhesion and proliferation.
, Lin-Gibson, S.
and Simon Jr., C.
Measuring Cell Adhesion and Proliferation in Polymer Scaffolds by X-Ray Microcomputed Tomography, Biomaterials, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=900996
(Accessed June 2, 2023)