(122) Examining Neuronal Marker Expression and Sensitivity to NSC348884 in a 3-D Multicellular Tumor Spheroid Neuroblastoma Model

Background: Neuroblastoma is an extracranial pediatric solid tumor with fewer than 60% of high-risk neuroblastoma patients reaching 5 years of survival. One of the hallmarks of neuroblastoma is the heterogeneity of both cellular and clinical features, and effective targeted therapies remain limited. 3-D cell culture models offer improved structural complexity and cellular heterogeneity compared with 2-D monolayer models, translating into new capabilities for drug testing. Our lab recently generated 3-D spheroids using two neuroblastoma cell lines, NLF and SH-SY5Y. Once these spheroids surpass 100 µm in diameter, they are referred to as Multicellular Tumor Spheroids (MCTSs).

Objectives: The objectives are to morphologically examine 3-D Multicellular Tumor Spheroids (MCTSs) formed from the NLF neuroblastoma cell line and to compare the sensitivity of MCTSs to 2-D monolayer cells to the small molecule NSC348884.

Design/Methods: 3-D Multicellular Tumor Spheroids, or MCTSs, were generated in serum-free media with B27 and EGF from the neuroblastoma cell line NLF. Immunofluorescence imaging was used to detect mature neuronal cell MAP2 and immature neuronal cell marker SOX2 in both MCTSs and 2-D monolayers. Both 2-D and 3-D cells were treated with 0-10 µM NSC348884 to measure drug sensitivity. 2-D drug sensitivity was measured using a colony formation assay. 3-D cell drug sensitivity was measured using spheroid diameter. Preliminary data indicate that MCTSs generated from NLF cells are more resistant to NSC348884 than 2-D monolayer cultures of NLF cells. Continued optimization and drug sensitivity testing in MCTSs are vital in improving the effectiveness, reproducibility, and thus scalability of this 3-dimensional MCTS neuroblastoma model.

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