This review was conducted to present the main neuroblastoma (NB) clinical characteristics and the most common genetic alterations present in these pediatric tumors, highlighting their impact in tumor cell aggressiveness behavior, including metastatic development and treatment resistance, and patients’ prognosis. The distinct three NB cell lineage phenotypes, S-type, N-type, and I-type, which are characterized by unique cell surface markers and gene expression patterns, are also reviewed. Finally, an overview of the most used NB cell lines currently available for in vitro studies and their unique cellular and molecular characteristics, which should be taken into account for the selection of the most appropriate model for NB pre-clinical studies, is presented. These valuable models can be complemented by the generation of NB reprogrammed tumor cells or organoids, derived directly from patients’ tumor specimens, in the direction toward personalized medicine. Impact statement This review provides an update on the mostly used cell line in vitro models for neuroblastoma (NB), a heterogeneous disease with high metastatic potential and resistance to treatment. The genetic and phenotypic profiles of the most used NB cell lines in the last 10 years are presented, considering the molecular markers that are involved in the distinct NB tumor phenotypes, including distinct core regulatory circuitries and non-coding RNAs. This gathered information can assist in the selection of the most appropriate NB in vitro model, based on the specific goals and objectives of each study.