Three-Dimensional Printing of Cell Exclusion Spacers (CES) for Use in Motility Assays
Cell migration/invasion assays are widely used in commercial drug discovery screening. 3D printing enables the creation of diverse geometric restrictive barrier designs for use in cell motility studies, permitting on-demand assays. Here, the utility of 3D printed cell exclusion spacers (CES) was validated as a cell motility assay.
A novel CES fit was fabricated using 3D printing and customized to the size and contour of 12 cell culture plates including 6 well plates of basal human brain vascular endothelial (D3) cell migration cells compared with 6 well plates with D3 cells challenged with 1uM cytochalasin D (Cyto-D), an F-actin anti-motility drug. Control and Cyto-D treated cells were monitored over 3 days under optical microscopy.
Day 3 cell migration distance for untreated D3 cells was 1515.943μm ± 10.346μm compared to 356.909μm ± 38.562μm for the Cyt-D treated D3 cells (p < 0.0001). By day 3, untreated D3 cells reached confluency and completely filled the original voided spacer regions, while the Cyt-D treated D3 cells remained significantly less motile.
Cell migration distances were significantly reduced by Cyto-D, supporting the use of 3D printing for cell exclusion assays. 3D printed CES have great potential for studying cell motility, migration/invasion, and complex multi-cell interactions.
Key words3D printing invasion assays migration assays motility assays personalized medicine three-dimensional printing
Cell exclusion spacers
- D3 cells
Human brain vascular endothelial cells
Food and drug administration
Migration and invasion
Phosphate-buffered saline/ethylenediaminetetraacetic acid
Acknowledgments and Disclosures
The authors would like to thank Louisiana State University Health Sciences Center Shreveport for supporting this research. The authors have no conflicts of interest to disclose.
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