A Human Glioblastoma Organotypic Slice Culture Model for Study of Tumor Cell Migration and Patient-specific Effects of Anti-Invasive Drugs

Abstract

Glioblastoma (GBM) continues to carry an extremely poor clinical prognosis despite surgical, chemotherapeutic, and radiation therapy. Progressive tumor invasion into surrounding brain parenchyma represents an enduring therapeutic challenge. To develop anti-migration therapies for GBM, model systems that provide a physiologically relevant background for controlled experimentation are essential. Here, we present a protocol for generating slice cultures from human GBM tissue obtained during surgical resection. These cultures allow for ex vivo experimentation without passaging through animal xenografts or single cell cultures. Further, we describe the use of time-lapse laser scanning confocal microscopy in conjunction with cell tracking to quantitatively study the migratory behavior of tumor cells and associated response to therapeutics. Slices are reproducibly generated within 90 min of surgical tissue acquisition. Retrovirally-mediated fluorescent cell labeling, confocal imaging, and tumor cell migration analyses are subsequently completed within two weeks of culture. We have successfully used these slice cultures to uncover genetic factors associated with increased migratory behavior in human GBM. Further, we have validated the model's ability to detect patient-specific variation in response to anti-migration therapies. Moving forward, human GBM slice cultures are an attractive platform for rapid ex vivo assessment of tumor sensitivity to therapeutic agents, in order to advance personalized neuro-oncologic therapy.

Multiplexed drug testing of tumor slices using a microfluidic platform

“…Additional phenotypic or response readouts include measures of signaling pathways 44 ; the induction of DNA damage responses (γH2AX staining) 8,22 ; cell proliferation (Ki-67 staining, Fig. S1); and cell migration and invasion 45 . All of these measures of intact tumor tissue response can be further extended by the analysis of slice culture tissue by biochemical 43,44 , flow cytometric 32 , and genomic approaches 44 .…”

The role of integrin αvand CD44 in GBM migration in human brain

“…Several groups have made use of human brain tissue in organotypic slice experimentation, focused on utilizing tumor tissue instead of normal brain [58,59]. A recent report presented a method for agarose embedding of tissue slices for cutting by a horizontal, vibratome blade [59].…”

Biological models in multiple sclerosis

“…Specific conditions in the organotypic slice cultures enable myelination of CNS tissue. Documented in various species; such as in mice (Huang et al, 2011;Mi et al, 2009), rats (Birgbauer, Rao, & Webb, 2004;Gianinazzi et al, 2005), and humans (Parker, Lizarraga, Waziri, & Foshay, 2017;Verwer et al, 2002), brain slice culture protocols allow the preservation of different cell types, structural aspects, synaptic organization, and electrical and biochemical pathways (Humpel, 2015). CNS organotypic cultures can be prepared from several spinal cord (Liu, Huang, Xiang, Zhao, & Huang, 2017) and brain regions, including cerebellum (Doussau et al, 2017), forebrain (Mi et al, 2009), and hippocampus (Gimsa, Peter, Lehmann, Bechmann, & Nitsch, 2000).…”