Primary Cell Culture of Central Neurocytomas

Lee, Seung Joon; Kim, Jeong Eun; Paek, Sun Ha; Keyoung, Hansoo Michael; Kim, Dong Gyu; Jung, Hee Won

Abstract

OBJECTIVE
The authors examine the characteristics of primary cell culture of central neurocytomas to discern the clues concerning the tumor origin. METHODS: Tumor cells of central neurocytomas from nine patients were cultured in 10% Dulbeco's Modified Essential Medium(DMEM)(eight) and in M21 media including basic Fibroblast Growth Factor(bFGF) and Epidermal Growth Factor(EGF)(one). The cultured cells had been stored in liquid nitrogen at the end of each passage, which were thawed and subcultured in one to three months. Morphological changes were chron-ologically examined under a phase contrast microscope. Immunocytochemical(ICC) stainings and Electron Microscopic(EM) examinations were performed in the early and late phases of the cultures to characterize the biological properties of the cultured cells. RESULTS: Within one to three days of primary culture, sprouting of cytoplasmic processes was observed with the size of the cells and the cytoplasmic processes increased. The cells stored from the liquid nitrogen showed the similar morphology to their original one before the storage within two to three days after thawing. ICC stainings of the cells cultured in 10% DMEM demonstrated dual differentiation. The cultured cells were positive for neuronal markers in the early stages and gial markers in the late stages. An EM study demonstrated both neuronal and glial differentiation regardless of the culture stages. The cells cultured in M21 including bFGF & EGF generated neurospheres and expressed the early neuronal protein, BIII-tubulin and a glial marker, GFAP. CONCLUSION: The central neurocytoma might therefore be a neoplasm of ventricular zone neural stem cells, multipotential when removed in vitro.