Morphology and growth, tumorigenicity, and cytogenetics of human neuroblastoma cells in continuous culture.

TLDR

Continuous cell lines SK‑N‑SH and SK‑N‑MC were established from human metastatic neuroblastoma tissue and maintained in vitro for 1–2 years. The SK‑N‑SH line contains spiny and epithelioid cells with high dopamine‑β‑hydroxylase activity, whereas SK‑N‑MC consists of fibroblast‑like cells lacking this activity but showing intracellular catecholamine fluorescence; both lines grow disoriented, reach high saturation densities, have rapid doubling times (44 h and 32 h), form tumors in 30–40 % of hamster cheek pouches, and are near‑diploid with characteristic chromosomal markers including double‑minute chromosomes.

Abstract

Summary Continuous cell lines, SK-N-SH and SK-N-MC, were established in cell culture from human metastatic neuroblastoma tissue and maintained in vitro for 1 to 2 years. SK-N-SH comprises two morphologically distinctive cell types, a small spiny cell and a large epithelioid cell. SK-N-MC is composed of small fibroblast-like cells with scant cytoplasm. In monolayer culture both cell lines form disoriented growth patterns and reach high saturation densities. Population-doubling times were 44 and 32 hr for SK-N-SH and SK-N-MC, respectively. Inoculum levels of 10 7 cells of both lines produced tumors confirmed by histopathological examination, at frequencies of 30 to 40% in cheek pouches of conditioned Syrian hamsters. SK-N-SH cells are characterized by high dopamine-β-hydroxylase activity while SK-N-MC cells have no detectable activity. However, for SK-N-MC but not SK-N-SH, the presence of intracellular catecholamine was indicated by formaldehyde-induced fluorescence. The lines are near-diploid with several chromosomal markers; SK-N-MC cells contain double- minute chromosomes. Growth, biochemical, and cytogenetic properties confirmed that the lines comprise malignant cells of neurogenic origin.