Breast Cancer Research and Treatment

, Volume 144, Issue 1, pp 93–101 | Cite as

Ex vivo Evans blue assessment of the blood brain barrier in three breast cancer brain metastasis models

  • John Do
  • Deshka Foster
  • Corinne Renier
  • Hannes Vogel
  • Sahar Rosenblum
  • Timothy C. Doyle
  • Victor Tse
  • Irene WapnirEmail author
Preclinical study


The limited entry of anticancer drugs into the central nervous system represents a special therapeutic challenge for patients with brain metastases and is primarily due to the blood brain barrier (BBB). Albumin-bound Evans blue (EB) dye is too large to cross the BBB but can grossly stain tissue blue when the BBB is disrupted. The course of tumor development and the integrity of the BBB were studied in three preclinical breast cancer brain metastasis (BCBM) models. A luciferase-transduced braintropic clone of MDA-231 cell line was used. Nude mice were subjected to stereotactic intracerebral inoculation, mammary fat pad-derived tumor fragment implantation, or carotid artery injections. EB was injected 30 min prior to euthanasia at various timepoints for each of the BCBM model animals. Serial bioluminescent imaging demonstrated exponential tumor growth in all models. Carotid BCBM appeared as diffuse multifocal cell clusters. EB aided the localization of metastases ex vivo. Tumor implants stained blue at 7 days whereas gross staining was not evident until day 14 in the stereotactic model and day 28 for the carotid model. EB assessment of the integrity of the BBB provides useful information relevant to drug testing in preclinical BCBM models.


Breast cancer brain metastasis Mouse brain metastasis model Blood brain barrier 



We would like to thank Dr. Sanjiv Sam Gambhir for his technical support. This study was supported in part by the Komen for the Cure grant (KG090545), Small Animal Imaging Resource NIH-NCI ICMIC P50-CA114747-02 (Gambhir, PI), and Stanford University Cancer Center NIH NCI CCSG P30-CA124435-02 (Mitchell, PI).


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • John Do
    • 1
  • Deshka Foster
    • 1
  • Corinne Renier
    • 1
  • Hannes Vogel
    • 2
  • Sahar Rosenblum
    • 3
  • Timothy C. Doyle
    • 4
  • Victor Tse
    • 3
  • Irene Wapnir
    • 1
    Email author
  1. 1.Department of SurgeryStanford University School of MedicineStanfordUSA
  2. 2.Department of PathologyStanford University School of MedicineStanfordUSA
  3. 3.Department of NeurosurgeryStanford University School of MedicineStanfordUSA
  4. 4.Department of PediatricsStanford University School of MedicineStanfordUSA

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