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Culturing Tumor and Normal Cells

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Dissociated cells (tumor-derived and normal NSCs) were plated at 50 000 cells/well in 6 well plates (Nunc, VWR Norge) coated with 15 μg/ml poly-L-ornithine (Sigma, St. Louis, MO). DMEM/F12 medium was augmented with fetal calf serum (3.75 %; PAA Laboratories, Pasching, Austria) and 25 μl/ml B-27 with vitamin A (Invitrogen), 1 μg/ml laminin and 10 μl/ml of Pen/Strep. All cultures were kept in an incubator at 5–6 % CO₂, pH 7.2-7.4 [40 (link)].

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Joel M., Mughal A.A., Grieg Z., Murrell W., Palmero S., Mikkelsen B., Fjerdingstad H.B., Sandberg C.J., Behnan J., Glover J.C., Langmoen I.A, & Stangeland B. (2015). Targeting PBK/TOPK decreases growth and survival of glioma initiating cells in vitro and attenuates tumor growth in vivo. Molecular Cancer, 14, 121.

Publication 2015

poly-L-ornithine fetal calf serum B-27 with vitamin A

Cells Fetal calf serum Laminin Poly l ornithine Strep Tumor Vitamin a

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Variable analysis

independent variables

Cell type (tumor-derived cells and normal NSCs)

dependent variables

Cell growth and proliferation

control variables

Seeding density (50,000 cells/well)
Culture substrate (6-well plates coated with 15 μg/ml poly-L-ornithine)
Culture medium (DMEM/F12 with 3.75% fetal calf serum, 25 μl/ml B-27 with vitamin A, 1 μg/ml laminin, and 10 μl/ml Pen/Strep)
Incubation conditions (5-6% CO2, pH 7.2-7.4)

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