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n2:pmcid: PMC0
bibo:doi: 10.1186%2F1471-2407-9-465
n5:contains: _:vb6706293 _:vb6706295 _:vb6706299 _:vb6706324

Subject Item: _:vb6706293
rdf:type: n5:Section
dc:title: results
n5:contains: _:vb6706294

Subject Item: _:vb6706294
rdf:type: n2:Context
rdf:value: The biopsy tissue was cut into 0.3 mm sized fragments that were cultured in vitro in agar-overlay cultures as previously described [>>14<<] and implanted as soon as spheroids had formed.
n2:mentions: n3:2406382

Subject Item: _:vb6706295
rdf:type: n5:Section
dc:title: methods
n5:contains: _:vb6706296 _:vb6706297 _:vb6706298

Subject Item: _:vb6706296
rdf:type: n2:Context
rdf:value: The study was approved by the regional Ethical Board at Haukeland University Hospital. Spheroids were prepared as described previously [>>14<<].
n2:mentions: n3:2406382

Subject Item: _:vb6706297
rdf:type: n2:Context
rdf:value: The athymic nude rat is T-cell deficient, but has normal complement and B-cell function [>>11<<]. 10 tumour spheroids (250-350 μm in diameter) were selected under a light microscope. The animals were anaesthetized with Hypnorm-Dormicum (0.4 ml/kg) s.c., the head secured in a stereotactic frame (Benchmark; Neurolab, St Louis, MO) and
n2:mentions: n3:307688

Subject Item: _:vb6706298
rdf:type: n2:Context
rdf:value: Animals engrafted with tissue from the initial 9 biopsies were imaged, using the same MRI combined with a small circular finger coil [>>21<<]. Subsequently, animal images were obtained, using a Bruker Pharmascan 7T MR scanner (Bruker Biospin MRI GmbH, Ettlingen, Germany), an axial T1 weighted MSME sequence (TR 1000 ms, TE 8.7 ms, slice thickness 1 mm, FOV 3.5 cm, matrix size
n2:mentions: n3:12892228

Subject Item: _:vb6706299
rdf:type: n5:Section
dc:title: discussion
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Subject Item: _:vb6706300
rdf:type: n2:Context
rdf:value: Outcome and treatment has improved only marginally over the last decades for GBMs [>>22<<], and reflect a limited understanding of the mechanisms that regulate their biological behaviour.
n2:mentions: n3:17618441

Subject Item: _:vb6706301
rdf:type: n2:Context
rdf:value: Transgenic brain tumour models, involve germline or somatic genetic modifications that predispose animals to developing CNS tumours [>>23<<-25]. However, whereas these models have a limited number of defined genetic changes, human brain tumours are characterised by multiple genetic aberrations. Alternatively, tumour development can be induced by carcinogen exposure or
n2:mentions: n3:16885338 n3:9851974 n3:10802656

Subject Item: _:vb6706302
rdf:type: n2:Context
rdf:value: Alternatively, tumour development can be induced by carcinogen exposure or established by implantation of [>>26<<,27] or xenogeneic [26] cancer cells or tumour biopsies [19,20].
n2:mentions: n3:17874037

Subject Item: _:vb6706303
rdf:type: n2:Context
rdf:value: Alternatively, tumour development can be induced by carcinogen exposure or established by implantation of [26,>>27<<] or xenogeneic [26] cancer cells or tumour biopsies [19,20].
n2:mentions: n3:7760095

Subject Item: _:vb6706304
rdf:type: n2:Context
rdf:value: Alternatively, tumour development can be induced by carcinogen exposure or established by implantation of [26,27] or xenogeneic [>>26<<] cancer cells or tumour biopsies [19,20].
n2:mentions: n3:17874037

Subject Item: _:vb6706305
rdf:type: n2:Context
rdf:value: Alternatively, tumour development can be induced by carcinogen exposure or established by implantation of [26,27] or xenogeneic [26] cancer cells or tumour biopsies [>>19<<,20]. While cell line based models often fail to mimic essential characteristics of brain tumours [4], some studies have reported growth of highly infiltrative tumours, following the implantation of human glioma biopsies in the nude rat
n2:mentions: n3:10413165

Subject Item: _:vb6706306
rdf:type: n2:Context
rdf:value: Alternatively, tumour development can be induced by carcinogen exposure or established by implantation of [26,27] or xenogeneic [26] cancer cells or tumour biopsies [19,>>20<<]. While cell line based models often fail to mimic essential characteristics of brain tumours [4], some studies have reported growth of highly infiltrative tumours, following the implantation of human glioma biopsies in the nude rat brain
n2:mentions: n3:17056721

Subject Item: _:vb6706307
rdf:type: n2:Context
rdf:value: While cell line based models often fail to mimic essential characteristics of brain tumours [>>4<<], some studies have reported growth of highly infiltrative tumours, following the implantation of human glioma biopsies in the nude rat brain [4,19,20].
n2:mentions: n3:12471461

Subject Item: _:vb6706308
rdf:type: n2:Context
rdf:value: While cell line based models often fail to mimic essential characteristics of brain tumours [4], some studies have reported growth of highly infiltrative tumours, following the implantation of human glioma biopsies in the nude rat brain [>>4<<,19,20]. Sakaria et al reported that the parent tumour's EGFR amplification status was preserved through subcutaneous passaging of the glioblastoma xenografts in nude mice [28].
n2:mentions: n3:12471461

Subject Item: _:vb6706309
rdf:type: n2:Context
rdf:value: cell line based models often fail to mimic essential characteristics of brain tumours [4], some studies have reported growth of highly infiltrative tumours, following the implantation of human glioma biopsies in the nude rat brain [4,>>19<<,20]. Sakaria et al reported that the parent tumour's EGFR amplification status was preserved through subcutaneous passaging of the glioblastoma xenografts in nude mice [28].
n2:mentions: n3:10413165

Subject Item: _:vb6706310
rdf:type: n2:Context
rdf:value: cell line based models often fail to mimic essential characteristics of brain tumours [4], some studies have reported growth of highly infiltrative tumours, following the implantation of human glioma biopsies in the nude rat brain [4,19,>>20<<]. Sakaria et al reported that the parent tumour's EGFR amplification status was preserved through subcutaneous passaging of the glioblastoma xenografts in nude mice [28].
n2:mentions: n3:17056721

Subject Item: _:vb6706311
rdf:type: n2:Context
rdf:value: Sakaria et al reported that the parent tumour's EGFR amplification status was preserved through subcutaneous passaging of the glioblastoma xenografts in nude mice [>>28<<]. Moreover, cell suspensions established from short-term in vitro cultures of flank xenograft tumours could subsequently be xenografted orthotopically. They successfully applied this approach to investigate how EGFR amplification status
n2:mentions: n3:16609043

Subject Item: _:vb6706312
rdf:type: n2:Context
rdf:value: Previously, we have also shown that first generation tumours derived from GBM biopsies often exhibit angiogenesis-independent growth while passaging is accompanied by the onset of angiogenesis, producing highly vascular tumours [>>20<<,29,30]. Moreover, gene expression profiling and protein arrays demonstrated that this phenotypic shift coincided with alterations in signalling pathways [20]. Whereas components of the Wnt, PI3K, and NF-kβ signalling pathways were
n2:mentions: n3:17056721

Subject Item: _:vb6706313
rdf:type: n2:Context
rdf:value: Previously, we have also shown that first generation tumours derived from GBM biopsies often exhibit angiogenesis-independent growth while passaging is accompanied by the onset of angiogenesis, producing highly vascular tumours [20,>>29<<,30]. Moreover, gene expression profiling and protein arrays demonstrated that this phenotypic shift coincided with alterations in signalling pathways [20].
n2:mentions: n3:17047051

Subject Item: _:vb6706314
rdf:type: n2:Context
rdf:value: Previously, we have also shown that first generation tumours derived from GBM biopsies often exhibit angiogenesis-independent growth while passaging is accompanied by the onset of angiogenesis, producing highly vascular tumours [20,29,>>30<<]. Moreover, gene expression profiling and protein arrays demonstrated that this phenotypic shift coincided with alterations in signalling pathways [20].
n2:mentions: n3:17955491

Subject Item: _:vb6706315
rdf:type: n2:Context
rdf:value: Moreover, gene expression profiling and protein arrays demonstrated that this phenotypic shift coincided with alterations in signalling pathways [>>20<<]. Whereas components of the Wnt, PI3K, and NF-kβ signalling pathways were overexpressed in the invasive first-generation tumours compared with the high-generation tumours, the Ras signalling pathway was up-regulated in the high generation
n2:mentions: n3:17056721

Subject Item: _:vb6706316
rdf:type: n2:Context
rdf:value: In our series, average survival was 118 days, whereas the average standard error of the mean (SEM) was 9.3 days, which compares well to other cell line based studies [>>31<<-33]. Moreover, 24 of 25 (96%) GBM biopsies produced tumours in 137 of 171 (80%) nude rats. These findings are similar to a previous smaller study in which 5 of 6 (83%) of the GBM biopsies produced tumours in 38 of 47 (81%) nude rats [19].
n2:mentions: n3:16278418 n3:11753988 n3:19487894

Subject Item: _:vb6706317
rdf:type: n2:Context
rdf:value: These findings are similar to a previous smaller study in which 5 of 6 (83%) of the GBM biopsies produced tumours in 38 of 47 (81%) nude rats [>>19<<]. This is also in line with numerous studies using primary biopsies from other subtypes that provide high tumour take rates [3,15-18,34-37]. The low take rates we observed with biopsies from grade II and III tumours may be due to a lack
n2:mentions: n3:10413165

Subject Item: _:vb6706318
rdf:type: n2:Context
rdf:value: This is also in line with numerous studies using primary biopsies from other subtypes that provide high tumour take rates [>>3<<,15-18,34-37].
n2:mentions: n3:10700174

Subject Item: _:vb6706319
rdf:type: n2:Context
rdf:value: This is also in line with numerous studies using primary biopsies from other subtypes that provide high tumour take rates [3,>>15<<-18,34-37].
n2:mentions: n3:1959996 n3:1332576 n3:1608975 n3:8517640

Subject Item: _:vb6706320
rdf:type: n2:Context
rdf:value: This is also in line with numerous studies using primary biopsies from other subtypes that provide high tumour take rates [3,15-18,>>34<<-37]. The low take rates we observed with biopsies from grade II and III tumours may be due to a lack of adaptability to the rat brain environment. However, while grade II gliomas may appear radiologically unchanged for several years in
n2:mentions: n3:1924398 n3:8352558 n3:8439965 n3:1444197

Subject Item: _:vb6706321
rdf:type: n2:Context
rdf:value: However, while grade II gliomas may appear radiologically unchanged for several years in the patient, the life span of nude rats is in the range of 1,5 -2 years under specific pathogen free conditions [>>38<<]. Therefore, the low take rates may simply reflect that the time required to develop clinically manifest tumours from low grade glioma biopsies exceeded the life span of the hosts. Although the number of lower grade tumours was small in
n2:mentions: n3:1606201

Subject Item: _:vb6706322
rdf:type: n2:Context
rdf:value: While we previously demonstrated that passaging the tumours for 4-5 generations was accompanied by an onset of angiogenesis [>>20<<], this study includes biopsies that have been passaged for up to 19 generations.
n2:mentions: n3:17056721

Subject Item: _:vb6706323
rdf:type: n2:Context
rdf:value: Using the same strategy on invasive and non-invasive tumours can help to identify invasion-related genes and proteins [>>29<<]. We therefore believe this model provides a powerful tool do dissect mechanisms that mediate angiogenesis and invasive growth.
n2:mentions: n3:17047051

Subject Item: _:vb6706324
rdf:type: n5:Section
dc:title: background
n5:contains: _:vb6706336 _:vb6706337 _:vb6706325 _:vb6706326 _:vb6706327 _:vb6706332 _:vb6706333 _:vb6706334 _:vb6706335 _:vb6706328 _:vb6706329 _:vb6706330 _:vb6706331

Subject Item: _:vb6706325
rdf:type: n2:Context
rdf:value: In tumour biology, model systems typically involve cancer cells that have been passaged numerous times in vitro to provide cancer cell lines [>>1<<]. In animals, the cells are often implanted subcutaneously, or in other ectopic locations [2] to produce experimental tumours. The fact that these models are easily established and standardized, favour their use in high throughput
n2:mentions: n3:4313504

Subject Item: _:vb6706326
rdf:type: n2:Context
rdf:value: In animals, the cells are often implanted subcutaneously, or in other ectopic locations [>>2<<] to produce experimental tumours.
n2:mentions: n3:6484200

Subject Item: _:vb6706327
rdf:type: n2:Context
rdf:value: Although cell line models provide highly reproducible results, these findings may have a limited clinical relevance [>>3<<,4]. Notably, only a fraction of the drug candidates that show promising results in animal studies is ever implemented in treatment protocols for cancer patients [5-8]. Thus, more powerful tools than the models that are currently used are
n2:mentions: n3:10700174

Subject Item: _:vb6706328
rdf:type: n2:Context
rdf:value: Although cell line models provide highly reproducible results, these findings may have a limited clinical relevance [3,>>4<<]. Notably, only a fraction of the drug candidates that show promising results in animal studies is ever implemented in treatment protocols for cancer patients [5-8]. Thus, more powerful tools than the models that are currently used are
n2:mentions: n3:12471461

Subject Item: _:vb6706329
rdf:type: n2:Context
rdf:value: Notably, only a fraction of the drug candidates that show promising results in animal studies is ever implemented in treatment protocols for cancer patients [>>5<<-8]. Thus, more powerful tools than the models that are currently used are needed.
n2:mentions: n3:11857365 n3:11135548 n3:14508090 n3:11559558

Subject Item: _:vb6706330
rdf:type: n2:Context
rdf:value: Orthotopic models are characterised by tumour growth in the animal, in a site corresponding to the human organ in which the tumour arises [>>9<<-11]. Therefore, such models provide an optimal system for studying tumour-host interactions, within an organ-specific niche [12,13].
n2:mentions: n3:307688 n3:1189920 n3:167965

Subject Item: _:vb6706331
rdf:type: n2:Context
rdf:value: Therefore, such models provide an optimal system for studying tumour-host interactions, within an organ-specific niche [>>12<<,13]. Moreover, engrafting primary tissue instead of cancer cell lines may prevent a genetic drift and phenotypic changes induced by prolonged passaging in vitro [14]. Indeed, orthotopic tumour models, using primary tissue, have been
n2:mentions: n3:16253523

Subject Item: _:vb6706332
rdf:type: n2:Context
rdf:value: Therefore, such models provide an optimal system for studying tumour-host interactions, within an organ-specific niche [12,>>13<<]. Moreover, engrafting primary tissue instead of cancer cell lines may prevent a genetic drift and phenotypic changes induced by prolonged passaging in vitro [14]. Indeed, orthotopic tumour models, using primary tissue, have been
n2:mentions: n3:18469852

Subject Item: _:vb6706333
rdf:type: n2:Context
rdf:value: Moreover, engrafting primary tissue instead of cancer cell lines may prevent a genetic drift and phenotypic changes induced by prolonged passaging in vitro [>>14<<]. Indeed, orthotopic tumour models, using primary tissue, have been established for several cancer subtypes [15-18], and studies have demonstrated that they can recapitulate tumour-host interactions and display traits of the parent tumour
n2:mentions: n3:2406382

Subject Item: _:vb6706334
rdf:type: n2:Context
rdf:value: Indeed, orthotopic tumour models, using primary tissue, have been established for several cancer subtypes [>>15<<-18], and studies have demonstrated that they can recapitulate tumour-host interactions and display traits of the parent tumour [19].
n2:mentions: n3:1959996 n3:1332576 n3:8517640 n3:1608975

Subject Item: _:vb6706335
rdf:type: n2:Context
rdf:value: orthotopic tumour models, using primary tissue, have been established for several cancer subtypes [15-18], and studies have demonstrated that they can recapitulate tumour-host interactions and display traits of the parent tumour [>>19<<].
n2:mentions: n3:10413165

Subject Item: _:vb6706336
rdf:type: n2:Context
rdf:value: Previously, we established an orthotopic biopsy-based model to study angiogenesis and malignant progression [>>20<<]. These experiments demonstrated that engraftment of GBM biopsies produced highly infiltrative gliomas that could grow in the absence of angiogenesis, and that passaging was accompanied by the onset of angiogenesis and shorter survival.
n2:mentions: n3:17056721

Subject Item: _:vb6706337
rdf:type: n2:Context
rdf:value: This series also included the xenograft tumours we previously used to investigate the invasive and vascular phenotypes in brain tumours [>>20<<]. However, the present work also provides experimental data about the growth parameters, neuropathological and radiological characteristics of these tumours.
n2:mentions: n3:17056721

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