Quantification of glioma cell invasion by confocal laser scanning microscopy in an in vitro co-culture system

Nygaard, Svein Jacob Tjoflaat; Tysnes, Ole‐Bjørn

doi:10.1016/0304-3835(96)04261-9

Cited by 21 publications

(20 citation statements)

References 13 publications

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“…The results obtained using fetal human and newborn rat astrocytes aggregates are consistent with data obtained for confrontation cultures of spheroid/fetal rat brain aggregates on agar (Bjerkvig et al, 1986). Invading glioma cells are not hindered by the presence of the newborn rat astrocyte aggregates and are able to infiltrate the normal fetal rat brain as demonstrated by scanning and confocal laser microscopy (Bjerkvig et al, 1986;Steinsvag, 1985;Nygaard et al, 1995;Engebraaten et al, 1990).…”

Section: Astrocyte Invasion and Migrationmentioning

confidence: 94%

Evidence for a secreted chemorepellent that directs glioma cell invasion

2004

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Secreted chemotropic cues guide the migration of neuronal and glial cell precursors during neural development. It is not known if chemotropism contributes to directing the invasion of brain tissue by glioma cells. A model system has been developed that allows quantification of invasive behavior using gliomas spheroids embedded in collagen gels. Here we provide evidence that glioma spheroids secrete a chemorepellent factor(s) that directs cells away from the spheroid and into the collagen matrix. The relationship between total invasion, cell number, and implantation distance suggests that glioma cells respond to a gradient of the chemorepellent cue(s) that is well established at 48 h. C6 astrocytoma cells normally invade the collagen at an angle perpendicular to the spheroid edge. In contrast, an adjacent spheroid causes cells to turn away from their normal trajectory and slow their rate of invasion. Astrocytoma cells are repelled by an adjacent glioma spheroid but rapidly infiltrate astrocyte aggregates, indicating that astrocytes do not express the repellent cue. Uniform concentrations of repellent factor(s) in spheroid conditioned medium overwhelm endogenous gradients and render glioma cells less able to exhibit this chemotropic response. Concentration gradients of spheroid conditioned medium in cell migration assays also demonstrate the chemorepellent cue(s)'s tropic effect. Our findings indicate that glioma spheroids produce a secreted diffusible cue(s) that promotes glioma cell invasion. Identification of this factor(s) may advance current therapies that aim to limit tumor cell invasion.

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Section: Astrocyte Invasion and Migrationmentioning

confidence: 94%

Evidence for a secreted chemorepellent that directs glioma cell invasion

2004

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“…[9][10][11] Moreover, confocal microscopy has been used to visualize glioma cell migration in rodent brain aggregates and organotypic brain slice models. [12][13][14][15] This body of work, and recent advances in organoid technology, 16 motivated the ex vivo development of a throughput-compatible biological assay. We based our methodology on neural tissue infiltration by free-floating human GBM and NP spheroids in co-culture with mouse embryonic stem cell (mESC)-derived eCOs.…”

Section: Introductionmentioning

confidence: 99%

Spontaneous Glioblastoma Spheroid Infiltration of Early-Stage Cerebral Organoids Models Brain Tumor Invasion

et al. 2018

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Organoid methodology provides a platform for the ex vivo investigation of the cellular and molecular mechanisms underlying brain development and disease. High-grade brain tumor glioblastoma multiforme (GBM) is considered a cancer of unmet clinical need, in part due to GBM cell infiltration into healthy brain parenchyma making complete surgical resection improbable. Modelling the process of GBM invasion in real time is challenging as it requires both tumor and neural tissue compartments. Here, we demonstrate that human GBM spheroids possess the ability to spontaneously infiltrate earlystage cerebral organoids (eCO). The resulting formation of hybrid organoids demonstrated an invasive tumor phenotype that was distinct from non-cancerous adult neural progenitor (NP) spheroid incorporation into eCOs. These findings provide a basis for the modelling and quantification of the GBM infiltration process using a stem cell-based organoid approach, and may be used for the identification of anti-GBM invasion strategies.

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“…In terms of a suitable matrix for the tumor, organotypic co-culture models have been used to study brain tumor invasion (Bjerkvig et al 1986;Bjerkvig et al 1990;Engebraaten et al 1990;Pedersen et al 1994a;Pedersen et al 1994b). Three-dimensional cultures involving two viable tissues confronting each other may accurately model the in vivo situation and provide more information than conventional membrane-based systems, where single tumor cells invade into a nonviable matrix (Ambrose and Easty 1973;Easty and Easty 1974;Nygaard et al 1995). Ohnishi et al (Ohnishi et al 1998), used rat brain slices obtained from the hippocampus or cortical regions of 2-day-old rats and maintained the brain slices in culture at the interface between air and the culture medium.…”

Section: Discussionmentioning

confidence: 99%