organism: Homo sapiens
attributes.tag: source_name
attributes.value: Tumour bulk cells from Mouse 1
attributes.tag: cell type
attributes.value: Patient-derived Glioma stem cell line (G144)
attributes.tag: host strain
attributes.value: CD-1 Nude (mouse)
attributes.tag: cell line
attributes.value: Patient derived line G144 expressing luciferase-GFP
attributes.tag: patient derived xenograft
attributes.value: 6-8 week old female CD-1 nude mice underwent stereotactic implantation of 1x105 GFP-luciferase expressing GSCs (anteroposterior 0, mediolateral -2.5, dorsoventral -3.5). Tumour growth was monitored using an IVIS Spectrum in vivo imaging system (Perkin Elmer). 10 minutes following i.p. D-luciferin (120 mg/kg, Intrace medical) bioluminescent images were acquired under isoflurane anaesthesia. Tumour size was quantified by calculating total flux (photons/s/cm2) using Living Image software (Xenogen, Caliper Life Sciences). Animals were sacrificed when they showed signs of distress or weight loss.
attributes.tag: facs sorting of tumour cells
attributes.value: Brains were harvested and tumour regions corresponding to tumour bulk, invaded striatum and corpus callosum were micro-dissected under fluorescent guidance in ice cold HEPES buffered HBSS. Micro-dissected regions were dissociated by incubating with papain (20 units/ml) DNase (0.005%) for 30min at 37°C (Worthington). Samples were triturated in HEPES buffered EBSS to fully dissociate the tissue and centrifuged (3 min, 300xg). Digestion was terminated by trituration in EBSS ovomucoid inhibitor (10mg/ml), albumin (10mg/ml) and DNase (0.005%). Following centrifugation (3 min, 300xg), cells were resuspended in 400 μL FACs sorting buffer containing 2.5% RNAsin and 1/10000 DAPI.
attributes.tag: culture conditions
attributes.value: NA
attributes.tag: treatment
attributes.value: NA