Perivascular invasion of primary human glioblastoma cells in organotypic human brain slices: human cells migrating in human brain

Rea Ravin; Paola Suarez-Meade; Brad Busse; Paul S Blank; Tito Vivas-Buitrago; Emily S Norton; Steve Graepel; Kaisorn L Chaichana; Ludmila Bezrukov; Hugo Guerrero-Cazares; Joshua Zimmerberg; Alfredo Quiñones-Hinojosa

doi:10.1007/s11060-023-04349-9

Perivascular invasion of primary human glioblastoma cells in organotypic human brain slices: human cells migrating in human brain

J Neurooncol. 2023 Aug;164(1):43-54. doi: 10.1007/s11060-023-04349-9. Epub 2023 Jul 25.

Authors

Affiliations

¹ Celoptics. Inc, Rockville, MD, USA.
² Department of Neurological Surgery, Mayo Clinic, Jacksonville, FL, USA.
³ Section On Integrative Biophysics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, 20892, USA.
⁴ Neuroscience Program, Mayo Clinic Graduate School of Biomedical Sciences, Jacksonville, USA.
⁵ Regenerative Sciences Training Program, Mayo Clinic Graduate School of Biomedical Sciences, Jacksonville, USA.
⁶ Section On Integrative Biophysics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, 20892, USA. zimmerberg.joshua@nih.gov.
⁷ Department of Neurological Surgery, Mayo Clinic, Jacksonville, FL, USA. quinones@mayo.edu.
⁸ Brain Tumor Stem Cell Laboratory, Department of Neurologic Surgery Mayo Clinic, 4500 San Pablo Rd S, Jacksonville, FL, 32224, USA. quinones@mayo.edu.

^# Contributed equally.

PMID: 37490233
DOI: 10.1007/s11060-023-04349-9

Abstract

Introduction: Glioblastoma (GBM) is an aggressive primary brain cancer. Lack of effective therapy is related to its highly invasive nature. GBM invasion has been studied with reductionist systems that do not fully recapitulate the cytoarchitecture of the brain. We describe a human-derived brain organotypic model to study the migratory properties of GBM IDH-wild type ex vivo.

Methods: Non-tumor brain samples were obtained from patients undergoing surgery (n = 7). Organotypic brain slices were prepared, and green fluorescent protein (GFP)-labeled primary human GBM IDH-wild type cells (GBM276, GBM612, GBM965) were placed on the organotypic slice. Migration was evaluated via microscopy and immunohistochemistry.

Results: After placement, cells migrated towards blood vessels; initially migrating with limited directionality, sending processes in different directions, and increasing their speed upon contact with the vessel. Once merged, migration speed decreased and continued to decrease with time (p < 0.001). After perivascular localization, migration is limited along the blood vessels in both directions. The percentage of cells that contact blood vessels and then continue to migrate along the vessel was 92.5% (- 3.9/ + 2.9)% while the percentage of cells that migrate along the blood vessel and leave was 7.5% (- 2.9/ + 3.9) (95% CI, Clopper-Pearson (exact); n = 256 cells from six organotypic cultures); these percentages are significantly different from the random (50%) null hypothesis (z = 13.6; p < 10^-7). Further, cells increase their speed in response to a decrease in oxygen tension from atmospheric normoxia (20% O₂) to anoxia (1% O₂) (p = 0.033).

Conclusion: Human organotypic models can accurately study cell migration ex vivo. GBM IDH-wild type cells migrate toward the perivascular space in blood vessels and their migratory parameters change once they contact vascular structures and under hypoxic conditions. This model allows the evaluation of GBM invasion, considering the human brain microenvironment when cells are removed from their native niche after surgery.

Keywords: Blood vessels; Brain-tumor initiating cells; Cell migration; Glioblastoma; Organotypic.

MeSH terms

Brain / pathology
Brain Neoplasms* / pathology
Cell Line, Tumor
Cell Movement / physiology
Glioblastoma* / pathology
Humans
Tumor Cells, Cultured
Tumor Microenvironment