Pharmaceutical Research

, 35:155 | Cite as

Three-Dimensional Printing of Cell Exclusion Spacers (CES) for Use in Motility Assays

  • Christen J. Boyer
  • David H. Ballard
  • Jungmi W. Yun
  • Adam Y. Xiao
  • Jeffery A. Weisman
  • Mansoureh Barzegar
  • Jonathan Steven AlexanderEmail author
Research Paper
Part of the following topical collections:
  1. 3D Printing of Pharmaceutical and Medical Applications: A New Era



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 words

3D printing invasion assays migration assays motility assays personalized medicine three-dimensional printing 



Cell exclusion spacers


Cytochalasin D

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Christen J. Boyer
    • 1
  • David H. Ballard
    • 2
  • Jungmi W. Yun
    • 1
  • Adam Y. Xiao
    • 1
  • Jeffery A. Weisman
    • 3
  • Mansoureh Barzegar
    • 1
  • Jonathan Steven Alexander
    • 1
    Email author
  1. 1.Molecular and Cellular PhysiologyLouisiana State University Health Sciences CenterShreveportUSA
  2. 2.Mallinckrodt Institute of RadiologyWashington University School of MedicineSt LouisUSA
  3. 3.Department of AnesthesiologyWashington University School of MedicineSt LouisUSA

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