Medulloblastoma tumor subtype identification

The SD-CSC and SI-CSC tumor subtype designation methods were reported previously (18 (link)). Briefly, freshly dissociated single cells were isolated from spontaneous medulloblastomas and plated at a low density (3,000 cells in 3 mL) in either TSC [modified DME/F-12 supplemented with B27 (Invitrogen) and penicillin/streptomycin] or NSC (TSC plus 20 ng/mL EGF and 10 ng/mL bFGF) medium in triplicates in 6-well plates, and secondary sphere formation was scored 5–7 days later. Tumors that formed secondary spheres only in NSC but not TSC medium were designated growth factor–dependent/SI-CSC subtype. Those that formed spheres in either NSC or TSC medium were designated as growth factor–independent/SD-CSC subtype. Those that did not form secondary spheres in either TSC or NSC medium were designated as no growth/SD-CSC subtype based on additional analyses (18 (link)).

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George J., Chen Y., Abdelfattah N., Yamamoto K., Gallup T.D., Adamson S.I., Rybinski B., Srivastava A., Kumar P., Lee M.G., Baskin D.S., Jiang W., Choi J.M., Flavahan W., Chuang J.H., Kim B.Y., Xu J., Jung S.Y, & Yun K. (2022). Cancer Stem Cells, not Bulk Tumor Cells, Determine Mechanisms of Resistance to SMO Inhibitors. Cancer Research Communications, 2(6), 402-416.

Publication 2022

DME/F-12

Cells Growth factor Medulloblastomas Penicillin Streptomycin Tumors

Corresponding Organization : Cornell University

Other organizations : Jackson Laboratory, The University of Texas MD Anderson Cancer Center, UConn Health, University of Maryland, Baltimore, University of Maryland Medical Center, The University of Texas Southwestern Medical Center, Baylor College of Medicine, University of Massachusetts Chan Medical School

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Variable analysis

independent variables

Cell culture medium (TSC or NSC)

dependent variables

Secondary sphere formation

control variables

Cell density (3,000 cells in 3 mL)
Plate format (6-well plates, triplicates)

controls

Positive control: Cells that formed secondary spheres in NSC medium (growth factor-dependent/SI-CSC subtype)
Negative control: Cells that did not form secondary spheres in either TSC or NSC medium (no growth/SD-CSC subtype)

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