Establishing MYCN-driven Tumor Cultures

Tumor tissue from mice were physically cut with scalpels into as small pieces as possible and dissociated with Accutase at 37°C for 10 min. Cells were then plated in 6-well plates in a modified version of NBE media^{40 (link)} consisting of Neurobasal media (Thermo Fisher), 1x Glutamax (Thermo Fisher), 1x N2 supplement (Thermo Fisher), 1x B27 without vitamin A supplement (Thermo Fisher), human recombinant FGF2 and EGF (both from Peprotech at 20 ng/mL). Doxycycline (Sigma Aldrich) was added at 1ug/mL to maintain expression of inducible MYCN. Media is changed every 2–3 days and tumor cells were passaged using Accutase.

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Huang M., Fang W., Farrel A., Li L., Chronopoulos A., Nasholm N., Cheng B., Zheng T., Yoda H., Barata M.J., Porras T., Miller M.L., Zhen Q., Ghiglieri L., McHenry L., Wang L., Asgharzadeh S., Park J., Gustafson W.C., Matthay K.K., Maris J.M, & Weiss W.A. (2024). ALK upregulates POSTN and WNT signaling to drive neuroblastoma. Cell reports, 43(3), 113927.

Publication 2024

Neurobasal media 1x Glutamax 1x N2 supplement human recombinant FGF2 EGF Doxycycline

Corresponding Organization : Neurological Surgery

Other organizations : Children's Hospital of Philadelphia, University of Pennsylvania

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Variable analysis

independent variables

Doxycycline (Sigma Aldrich) added at 1ug/mL to maintain expression of inducible MYCN

dependent variables

Tumor cell growth and proliferation

control variables

Tumor tissue from mice
Physical dissociation with scalpels
Dissociation with Accutase at 37°C for 10 min
Plating in 6-well plates
Modified version of NBE media consisting of Neurobasal media (Thermo Fisher), 1x Glutamax (Thermo Fisher), 1x N2 supplement (Thermo Fisher), 1x B27 without vitamin A supplement (Thermo Fisher), human recombinant FGF2 and EGF (both from Peprotech at 20 ng/mL)
Media change every 2–3 days
Tumor cell passaging using Accutase

controls

No positive or negative controls were explicitly mentioned in the provided information.

Annotations

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