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Cryo-image Analysis of Tumor Cell Migration, Invasion, and Dispersal in a Mouse Xenograft Model of Human Glioblastoma Multiforme

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Abstract

Purpose

The goals of this study were to create cryo-imaging methods to quantify characteristics (size, dispersal, and blood vessel density) of mouse orthotopic models of glioblastoma multiforme (GBM) and to enable studies of tumor biology, targeted imaging agents, and theranostic nanoparticles.

Procedures

Green fluorescent protein-labeled, human glioma LN-229 cells were implanted into mouse brain. At 20–38 days, cryo-imaging gave whole brain, 4-GB, 3D microscopic images of bright field anatomy, including vasculature, and fluorescent tumor. Image analysis/visualization methods were developed.

Results

Vessel visualization and segmentation methods successfully enabled analyses. The main tumor mass volume, the number of dispersed clusters, the number of cells/cluster, and the percent dispersed volume all increase with age of the tumor. Histograms of dispersal distance give a mean and median of 63 and 56 μm, respectively, averaged over all brains. Dispersal distance tends to increase with age of the tumors. Dispersal tends to occur along blood vessels. Blood vessel density did not appear to increase in and around the tumor with this cell line.

Conclusion

Cryo-imaging and software allow, for the first time, 3D, whole brain, microscopic characterization of a tumor from a particular cell line. LN-229 exhibits considerable dispersal along blood vessels, a characteristic of human tumors that limits treatment success.

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Acknowledgments

The authors would like to acknowledge the expert technical help of Miss Catherine Doller and Dr. Scott Howell with tissue histology. This work was supported by the Case Center for Imaging Research. The research was also supported by grants Ohio Wright Center/BRTT, The Biomedical Structure, Functional and Molecular Imaging Enterprise (D.L.W), National Institutes of Health grants R42CA124270 (D.L.W.), T32EB007509 (K.E.S), R01-NS051520 (S.B.-K.), and R01-NS063971 (S.B.-K., D.L.W. and J.P.B.).

Conflict of Interest

Debashish Roy, David L. Wilson, Mohammed Q. Qutaish, and Kristin E. Sullivant are inventors of cryo-imaging technology licensed by CWRU. Debashish Roy and David L. Wilson are officers with ownership positions at BioInVision, Inc., which is commercializing cryo-imaging.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Case Western Reserve University, Room 319 Wickenden Bldg., 2071 Martin Luther King Jr. Drive, Cleveland, OH, 44106-7207, USA

    Mohammed Q. Qutaish, Kristin E. Sullivant, Hong Lu, Debashish Roy, Jing Wang, James P. Basilion & David L. Wilson

  2. Department of Radiology, University Hospitals of Cleveland, Cleveland, OH, USA

    James P. Basilion & David L. Wilson

  3. Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH, USA

    Susan M. Burden-Gulley & Susann M. Brady-Kalnay

  4. Department of Neuroscience, Case Western Reserve University, Cleveland, OH, USA

    Susann M. Brady-Kalnay

  5. NFCR Center for Molecular Imaging, Case Western Reserve University, Cleveland, OH, USA

    James P. Basilion & David L. Wilson

Authors
  1. Mohammed Q. Qutaish
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  2. Kristin E. Sullivant
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  3. Susan M. Burden-Gulley
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  4. Hong Lu
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  5. Debashish Roy
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  6. Jing Wang
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  7. James P. Basilion
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  8. Susann M. Brady-Kalnay
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  9. David L. Wilson
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Corresponding author

Correspondence to David L. Wilson.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Whole brain visualization of blood vessels and tumor 1. 3D rendering of bright field volume of a 20-day-old tumor fades to show the results of the blood vessel enhancement algorithm (red) along with the results of the main tumor mass and dispersal cell detection algorithms (green and yellow, respectively). Rotation around the brain clearly shows the ability of the blood vessel enhancement algorithm to visualize blood vessels without the need for a contrast agent. (M4V 5814 kb)

LN-229 cell dispersal along blood vessels. ROI around LN-229 tumor at 20 days post-implantation shows blood vessels, main tumor mass, and dispersed cell clusters in red, green, and yellow, respectively. Dispersal along blood vessel is clearly shown by rotation and zooming (tumor 1). (M4V 5948 kb)

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Qutaish, M.Q., Sullivant, K.E., Burden-Gulley, S.M. et al. Cryo-image Analysis of Tumor Cell Migration, Invasion, and Dispersal in a Mouse Xenograft Model of Human Glioblastoma Multiforme. Mol Imaging Biol 14, 572–583 (2012). https://doi.org/10.1007/s11307-011-0525-z

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  • DOI: https://doi.org/10.1007/s11307-011-0525-z

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