Glioma Cells and Glioblastoma Stem Cell Culture

Human U87 glioma cells were obtained from American Type Culture Collection (ATCC, Manassas, VA) and grown in complete Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 % fetal calf serum at 37 °C and 5 % CO₂. Human GSCs MGG4, MGG8, MGG18, BT74 were isolated as previously described [28 (link),30 (link)], and maintained as spheres in serum-free medium containing 20 ng/mL recombinant human EGF (R&D systems) and 20 ng/mL recombinant human FGF2 (Peprotech). GSCs were passaged by dissociating neurospheres using the Neuro-Cult Chemical Dissociation Kit (StemCell Technologies). Mouse 005 GSCs were provided by Dr. I. Verma (Salk Institute for Biological Studies, La Jolla, CA) [33 (link), 34 (link)]. Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza. Human brain microvascular endothelial cells (HBMECs) were obtained from Dr. Ken Arai (MGH). Axitinib (Pfizer Inc) was provided by Pfizer and dissolved in DMSO as a 25 mM stock solution for in vitro studies. The final concentrations added to cells had less than 0.5 % DMSO, which was nontoxic to cells.

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Lu L., Saha D., Martuza R.L., Rabkin S.D, & Wakimoto H. (2014). Single agent efficacy of the VEGFR kinase inhibitor axitinib in preclinical models of glioblastoma. Journal of neuro-oncology, 121(1), 91-100.

Publication 2014

U87 glioma cells recombinant human EGF recombinant human FGF2 Neuro-Cult Chemical Dissociation Kit Human umbilical vein endothelial cells (HUVECs) Axitinib

Axitinib Biological Brain Cells Dmso Eagle Endothelial cells Fetal calf serum Glioma Human Human umbilical vein endothelial cells Mouse Recombinant human fgf2 Serum Stemcell

Corresponding Organization :

Other organizations : Massachusetts General Hospital, Harvard University

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Variable analysis

independent variables

Axitinib (Pfizer Inc) was provided by Pfizer and dissolved in DMSO as a 25 mM stock solution for in vitro studies. The final concentrations added to cells had less than 0.5 % DMSO, which was nontoxic to cells.

dependent variables

Not explicitly mentioned

control variables

Human U87 glioma cells were obtained from American Type Culture Collection (ATCC, Manassas, VA) and grown in complete Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal calf serum at 37 °C and 5 % CO2.
Human GSCs MGG4, MGG8, MGG18, BT74 were isolated as previously described [28 (link), 30 (link)], and maintained as spheres in serum-free medium containing 20 ng/mL recombinant human EGF (R&D systems) and 20 ng/mL recombinant human FGF2 (Peprotech).
Mouse 005 GSCs were provided by Dr. I. Verma (Salk Institute for Biological Studies, La Jolla, CA) [33 (link), 34 (link)].
Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza.
Human brain microvascular endothelial cells (HBMECs) were obtained from Dr. Ken Arai (MGH).

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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