Development human neuroblastoma cell line

The implementation of animal test protocols in the 1980s has been accompanied by the development of a host of alternative methods to study adverse effects of chemicals on reproductive and developmental parameters. For example, rat whole embryo culture stems from the seventies (16), as does the rat limb bud organ culture (17) and rat limb bud and brain micromass was developed in the eighties (18). An elegant nonvertebrate alternative model used regeneration of polyps of Hydra atUnuata from dissociated cells (19). Animal-free in vitro alternatives include those employing the proliferation of a human embryonic palatal mesenchymal cell line (20), the attachment of a mouse ovarian tumor cell line (21), and the differentiation of a neuroblastoma cell line (22) and a embryonal carcinoma cell line (23). Various overviews of methods have been published over the years (24). The predictability of... 

FIGURE 2.2 Methylation-specific PCR (MSP) analysis of the MDR-1 gene promoter methylation in human neuroblastoma (NB) cell lines (procured from Children s Oncology Group [COG] Philadelphia, PA) established from patients after chemotherapy.Methylation status of MDR-1 gene is represented as unmethylated (U) and methylated (M) in each cell type. A total of 19 neuroblastoma cell lines ° were examined by MSP and data indicate hypomethylation of MDR-1 gene in all cell lines tested. Hypomethylation (U) of MDR-1 gene indicates its increased expression and development of resistant phenotype in neuroblastoma cells after intensive chemotherapy. 

The simplest in vitro model, the cell line, is a population of cells that can be maintained in culture for an extended period of time. Neuronal cell lines normally originate from a single common ancestor cell (clonal) and are often derived from tumors, e.g., pha-eochromocytomas (adrenal medullary tumor) (e.g., PC12 cell line) [13] and neuroblastomas [14] such as mouse N2a or human SH-SY5Y. However, a recently developed technique to introduce oncogenes into primary cultures through retroviruses has opened up new possibilities [15]. 

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