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Development and characterization of a human orthotopic neuroblastoma xenograft.

Developmental biology (2015-04-12)
Elizabeth Stewart, Anang Shelat, Cori Bradley, Xiang Chen, Sara Federico, Suresh Thiagarajan, Abbas Shirinifard, Armita Bahrami, Alberto Pappo, Chunxu Qu, David Finkelstein, Andras Sablauer, Michael A Dyer
ABSTRACT

Neuroblastoma is a pediatric cancer of the developing sympathoadrenal lineage. The tumors are known to develop from the adrenal gland or paraspinal ganglia and have molecular and cellular features of sympathetic neurons such as dense core vesicles and catecholamine production. Here we present the detailed molecular, cellular, genetic and epigenetic characterization of an orthotopic xenograft derived from a high-risk stage 4 neuroblastoma patient. Overall, the xenografted tumor retained the high risk features of the primary tumor and showed aggressive growth and metastasis in the mouse. Also, the genome was preserved with no additional copy number variations, structural variations or aneuploidy. There were 13 missense mutations identified in the xenograft that were not present in the patient's primary tumor and there were no new nonsense mutations. None of the missense mutations acquired in the xenograft were in known cancer genes. We also demonstrate the feasibility of using the orthotopic neuroblastoma xenograft to test standard of care chemotherapy and molecular targeted therapeutics. Finally, we optimized a new approach to produce primary cultures of the neuroblastoma xenografts for high-throughput drug screening which can be used to test new combinations of therapeutic agents for neuroblastoma.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Deoxyribonuclease I from bovine pancreas, Type II-S, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein
Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture