Protocol detail

Glioma Cell Lines and Glioma Stem Cells

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The human glioma cell lines, LN229 and A172, were obtained from ATCC (Manassas, VA). U251HF glioma cells were kindly provided by Dr. W. K. Alfred Yung (M. D. Anderson Cancer Center, Houston, TX); U251HF-Luc were generated by stably transfecting a firefly luciferase gene under a CMV promoter into U251HF cells and maintained as described with the addition of Geneticin (G418–800μg/ml) for U251HF-Luc cells (23 (link)). GSC2, GSC20, GSC11, GSC23 and GSC811 patient-derived glioma stem cell lines (24 (link)) were kindly provided by Dr. Frederick Lang (M. D. Anderson Cancer Center, Houston, TX) and cultured as neurospheres passaged every 5–7 days in serum-free DMEM/F12 medium containing B-27 Supplement (Life Technologies, Carlsbad, CA), bFGF (FGF2 Gold BioTechnology, St. Luis, MO) and EGF (Gold Bio Technology). Cell lines were authenticated at the University of Arizona Genetics Core (http://uagc.arl.arizona.edu/services/cell-line-authentication-human). Onalespib (AT13387) was purchased from Medkoo Biosciences (Morrisville, NC).

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Canella A., Welker A.M., Yoo J.Y., Xu J., Abas F.S., Kesanakurti D., Nagarajan P., Beattie C.E., Sulman E.P., Liu J., Gumin J., Lang F.F., Gurcan M.N., Kaur B., Sampath D, & Puduvalli V.K. (2017). Efficacy of Onalespib a Long-acting Second Generation HSP90 Inhibitor as a Single Agent and in Combination with Temozolomide against Malignant Gliomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(20), 6215-6226.

Publication 2017

LN229 B-27 Supplement bFGF EGF

A gene At13387 Cancer Cell line authentication Cell lines Cells Fgf2 Firefly luciferase G418 Geneticin Glioma Gold Human Onalespib Patient Serum Stem Supplement

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Variable analysis

independent variables

Cell lines (LN229, A172, U251HF, U251HF-Luc, GSC2, GSC20, GSC11, GSC23, GSC811)

dependent variables

Not explicitly mentioned

control variables

Geneticin (G418–800μg/ml) for U251HF-Luc cells
Culturing conditions for patient-derived glioma stem cell lines (neurospheres passaged every 5–7 days in serum-free DMEM/F12 medium containing B-27 Supplement, bFGF, and EGF)

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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