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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 Alexander
Research Paper
Part of the following topical collections:
  1. 3D Printing of Pharmaceutical and Medical Applications: A New Era

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.

Key words

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

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

Notes

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.

References

  1. 1.
    Justus CR, Leffler N, Ruiz-Echevarria M, Yang LV. In vitro cell migration and invasion assays. J Vis Exp. 2014;(88) http://www.jove.com/video/51046/in-vitro-cell-migration-and-invasion-assays. Accessed March 22, 2018.
  2. 2.
    Kramer N, Walzl A, Unger C, Rosner M, Krupitza G, Hengstschläger M, et al. In vitro cell migration and invasion assays. Mutat Res Rev Mutat Res. 2013;752(1):10–24.CrossRefGoogle Scholar
  3. 3.
    Javaherian S, O’Donnell KA, McGuigan AP. A Fast and accessible methodology for micro-patterning cells on standard culture substrates using Parafilm™ inserts. Neves NM, editor. PLoS One. 2011;6(6):e20909.CrossRefGoogle Scholar
  4. 4.
    Weisman JA, Nicholson JC, Tappa K, Jammalamadaka U, Wilson CG, Mills DK. Antibiotic and chemotherapeutic enhanced three-dimensional printer filaments and constructs for biomedical applications. Int J Nanomedicine. 2015;10:357–70.PubMedPubMedCentralGoogle Scholar
  5. 5.
    Ballard DH, Weisman JA, Jammalamadaka U, Tappa K, Alexander JS, Griffen FD. Three-dimensional printing of bioactive hernia meshes: in vitro proof of principle. Surgery. 2017;161(6):1479–81.CrossRefGoogle Scholar
  6. 6.
    Weisman JA, Ballard DH, Jammalamadaka U, et al. 3D printed antibiotic and chemotherapeutic eluting catheters for potential use in interventional radiology: in vitro proof of concept study. Acad Radiol. 2018; [Ahead of print];  https://doi.org/10.1016/j.acra.2018.03.022.
  7. 7.
    Ballard DH, Trace AP, Ali S, Hodgdon T, Zygmont ME, DeBenedectis CM, et al. Clinical applications of 3D printing: primer for radiologists. Acad Radiol. 2018;25(1):52–65.CrossRefGoogle Scholar
  8. 8.
    Hodgdon T, Danrad R, Patel MJ, Smith SE, Richardson ML, Ballard DH, et al. Logistics of three-dimensional printing: primer for radiologists. Acad Radiol. 2018;25(1):40–51.CrossRefGoogle Scholar
  9. 9.
    Boyer CJ, Ballard DH, Weisman JA, Hurst S, McGee DJ, Mills DK, et al. Three-dimensional printing antimicrobial and radiopaque constructs. 3D Print Addit Manuf. 2018;5(1):29–35.CrossRefGoogle Scholar
  10. 10.
    Tappa K, Jammalamadaka U, Ballard DH, Bruno T, Israel MR, Vemula H, et al. Medication eluting devices for the field of OBGYN (MEDOBGYN): 3D printed biodegradable hormone eluting constructs, a proof of concept study. PLoS One. 2017;12(8):e0182929.CrossRefGoogle Scholar
  11. 11.
    Kang H-W, Lee SJ, Ko IK, Kengla C, Yoo JJ, Atala A. A 3D bioprinting system to produce human-scale tissue constructs with structural integrity. Nat Biotechnol. 2016;34(3):312–9.CrossRefGoogle Scholar
  12. 12.
    Laronda MM, Rutz AL, Xiao S, Whelan KA, Duncan FE, Roth EW, et al. A bioprosthetic ovary created using 3D printed microporous scaffolds restores ovarian function in sterilized mice. Nat Commun. 2017;8:15261.CrossRefGoogle Scholar
  13. 13.
    Casella JF, Flanagan MD, Lin S. Cytochalasin D inhibits actin polymerization and induces depolymerization of actin filaments formed during platelet shape change. Nature. 1981;293(5830):302–5.CrossRefGoogle Scholar
  14. 14.
    Di Prima M, Coburn J, Hwang D, et al. Additively manufactured medical products–the FDA perspective. 3D Print Med. 2016;2:1.CrossRefGoogle Scholar
  15. 15.
    Christensen A, Rybicki FJ. Maintaining safety and efficacy for 3D printing in medicine. 3D Printing in Medicine. 2017;3  https://doi.org/10.1186/s41205-016-0009-5.
  16. 16.
    Leng S, McGee K, Morris J, Alexander A, Kuhlmann J, Vrieze T, et al. Anatomic modeling using 3D printing: quality assurance and optimization. 3D Printing in Medicine. 2017;3  https://doi.org/10.1186/s41205-017-0014-3.
  17. 17.
    Morrison RJ, Kashlan KN, Flanangan CL, Wright JK, Green GE, Hollister SJ, et al. Regulatory considerations in the design and manufacturing of implantable 3d-printed medical devices. Clin Transl Sci. 2015;8(5):594–600.CrossRefGoogle Scholar

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
  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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