A rapid assay for drug sensitivity of glioblastoma stem cells.
Glioblastoma (GBM) is a highly infiltrating, aggressive brain cancer with no available curative treatment. We developed a rapid assay for assessing the effect of various drugs on GBM stem cells. The assay uses a small number of separated CD133+ cells (20,000 in 0.2 ml) in 96-well plate that form neurospheres within 1-2 days. Various drugs disperse the neurospheres within 24-36 h, which can be quantified microscopically. We used the GBM cell line A-172 to develop the conditions for the assay, utilizing Gleevec, the gamma-secretase inhibitor DAPT, and the anti-bacterial peptide amph1D. The results show dispersion of the neurospheres leading to cell death, at relatively low drugs concentrations (<25 microM). Drug combination showed a synergistic effect and disruption of neurospheres under lower concentrations. We applied this assay to the CD133+ cells of surgical specimens from three patients that showed similar results. This assay facilitates a rapid test of drugs on small amounts of fractionated patient's GBM stem cells.
Gal
Hilah
H
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
Makovitzki
Arik
A
Amariglio
Ninette
N
Rechavi
Gideon
G
Ram
Zvi
Z
Givol
David
D
D000071916:AC133 Antigen; D000890:Anti-Infective Agents / Q000494:pharmacology; D015703:Antigens, CD / Q000096:biosynthesis; D000970:Antineoplastic Agents / Q000494:pharmacology; D001549:Benzamides; D001932:Brain Neoplasms / Q000188:drug therapy*; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005909:Glioblastoma / Q000188:drug therapy*; D006023:Glycoproteins / Q000096:biosynthesis; D006801:Humans; D000068877:Imatinib Mesylate; D007150:Immunohistochemistry; D010455:Peptides / Q000737:chemistry; D010879:Piperazines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology; D013997:Time Factors
D000071916:AC133 Antigen; D000890:Anti-Infective Agents; D015703:Antigens, CD; D000970:Antineoplastic Agents; D001549:Benzamides; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D010879:Piperazines; D011743:Pyrimidines; D000068877:Imatinib Mesylate