Glioma Stem Cell Culture and Manipulation Protocol

GSC11, GSC23, GSC262, GSC811, GSC214, GSC231 and GSC267, GS2, GS272 and GS711 patient-derived glioma stem cell lines were cultured as neurospheres passaged every 4-7 days in serum-free DMEM/F12 medium containing B-27 Supplement (Life Technologies, Carlsbad, CA), EGF (Gold Bio Technology) and bFGF (FGF2 Gold BioTechnology, St. Louis, MO) (21 (link)). U251HF glioma cells were kindly provided by Dr. W. K. Alfred Yung (M. D. Anderson Cancer Center, Houston, TX). The generation and maintenance of the chemiluminescent U251HF-Luc and GSC811-Luc cells were as previously described (16 (link),22 (link)). Cell lines were authenticated at the University of Arizona Genetics Core (https://uagc.arl.arizona.edu/services/complete-solutions/cell-line-authentication). Onalespib (AT13387) was purchased from Medkoo Biosciences, Inc. (Morrisville, NC). Ionizing radiation was delivered alone or in combination with onalespib as described in figure legend.

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Xu J., Wu P.J., Lai T.H., Sharma P., Canella A., Welker A.M., Beattie C.E., Elder J.B., Easley M., Lonser R., Jacob N.K., Pietrzak M., Timmers C.M., Lang F., Sampath D, & Puduvalli V.K. (2022). Disruption of DNA Repair and Survival Pathways through Heat Shock Protein inhibition by Onalespib to Sensitize Malignant Gliomas to Chemoradiation therapy. Clinical cancer research : an official journal of the American Association for Cancer Research, 28(9), 1979-1990.

Publication 2022

B-27 Supplement EGF

At13387 Cancer Cell line authentication Cell lines Cells Fgf2 Glioma Gold Ionizing radiation Onalespib Patient Serum Stem Supplement

Corresponding Organization : The University of Texas MD Anderson Cancer Center

Other organizations : The Ohio State University, Medical University of South Carolina

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Variable analysis

independent variables

Onalespib (AT13387)
Ionizing radiation

dependent variables

Glioma stem cell line growth and characteristics

control variables

Serum-free DMEM/F12 medium containing B-27 Supplement, EGF, and bFGF
Cell line authentication

controls

Positive control: U251HF glioma cells
Negative control: Not explicitly mentioned

Annotations

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