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

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Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusions

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.

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Abbreviations

CES:

Cell exclusion spacers

Cyto-D:

Cytochalasin D

D3 cells:

Human brain vascular endothelial cells

FDA:

Food and drug administration

Migration/invasion:

Migration and invasion

PBS-EDTA:

Phosphate-buffered saline/ethylenediaminetetraacetic acid

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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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Authors and Affiliations

  1. Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA

    Christen J. Boyer, Jungmi W. Yun, Adam Y. Xiao, Mansoureh Barzegar & Jonathan Steven Alexander

  2. Mallinckrodt Institute of Radiology, Washington University School of Medicine, St Louis, Missouri, USA

    David H. Ballard

  3. Department of Anesthesiology, Washington University School of Medicine, St Louis, Missouri, USA

    Jeffery A. Weisman

Authors
  1. Christen J. Boyer
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  2. David H. Ballard
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  3. Jungmi W. Yun
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  4. Adam Y. Xiao
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  5. Jeffery A. Weisman
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  6. Mansoureh Barzegar
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  7. Jonathan Steven Alexander
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Corresponding author

Correspondence to Jonathan Steven Alexander.

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Guest Editors: Tony Zhou and Tonglei Li

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Boyer, C.J., Ballard, D.H., Yun, J.W. et al. Three-Dimensional Printing of Cell Exclusion Spacers (CES) for Use in Motility Assays. Pharm Res 35, 155 (2018). https://doi.org/10.1007/s11095-018-2431-4

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  • DOI: https://doi.org/10.1007/s11095-018-2431-4

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