Establishment of Korean Neuroblastoma Cell Line (NBL-K1) / 대한소아혈액종양학회지

ABSTRACT

PURPOSE: A new continuous cell line, NBL-K1, was established in tissue culture from a Korean child with stage IV neuroblastoma, arising from the adrenal gland, which had normal urinary excretion of VMA and HVA and diagnosed by light and electron microscope. METHODS: Clinical characteristics of patient was high ferritin level, normal neuron specific enolase, and normal urinary VMA and HVA. The small tissue specimen obtained from surgically resected tumor was minced with a mosquito scissors and scalpels and cultured in L-15 medium with 17% FBS (37oC and 5% CO2). Chromosome analysis was performed from bone marrow cell culture with a method of high resolution banding using methotrexate and thymidine and TGT staining. Chromosomes were analyzed by ISCN. The N-myc amplification was checked by N-myc primers, PCR, and gel electrophoresis. RESULTS: The cells were attached to the bottom of culture flask on 4th day of culture and composed of a small and elongated cell body with relatively abundant granules in cytoplasm and oval shaped nucleus with one prominent nucleoli and slender nerve-like fiber. Cell clumps were observed on 10th day of culture. The morphology was changed to round cell when trypsin was added. The chromosome analysis revealed two kinds of hyperdiploidy. No cell contained homogeneously stained region (HSR). But numerous double minutes (DMs) were observed. N- myc oncogene of the NBL-K1 was not amplified. The cultured cells with many black immunobeads around the surface considered to be the neuroblastoma cells. CONCLUSION: The characteristic Korean neuroblastoma cell line (NBL K-1) was estblished for the future studies of in vitro chemosensitivity test, monoclonal antibody and xenograft.