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Two new human neuroblastoma cell lines exhibiting tumorigenesis and metastasis in the nude mouse model
C L Fowler1, C C Zimmer, D Ruktanouchai
1Department of Surgery, University of Kentucky Medical Center, Lexington 40536, USA.
Background:
Human neuroblastoma cell lines are notoriously difficult to establish in culture and use in murine hosts.
Materials And Methods:
Two new human neuroblastoma cell lines, NK and ND, were established and studied for growth patterns in nude mice, growth in soft agar, cell cycle analysis, apoptosis (Hoechst- merocyanine 540 test), metalloproteinase expression (zymograms), and morphological differentiation by dibutyryl cyclic AMP (dCAMP).
Results:
Both cell lines formed tumors in 6/9 nude mice within 5-31 days after subcutaneous inoculation, and metastases after intravenous tail vein injection. Both grew in soft agar. DCAMP induced morphologic differentiation in both, and inhibited cell culture growth without apoptosis. Zymograms of supernatants from cultures revealed 72-kDa metalloproteinase and higher molecular bands that did not change with dCAMP treatment. Cultures derived from murine metastatic foci exhibited 72, 82 and 85-kDa proteins, with strong 92-kDa bands after dCAMP treatment.
Conclusion:
New human neuroblastoma cell lines were established that are easily used in nude mice, and express metalloproteinases.