HG-18 * A ROLE OF SPINDLE CELLS IN GLIOBLASTOMA GROWTH

Abstract

BACKGROUND: Bailey and Cushing believed gliomas originated from primitive spongioblasts (1926). The authors have found the spindle cells in the early stage of rat C6 cell cuture, resembling spongioblasts. Additionally, C6 cells implanted into rat brain grew glioblastoma (GBM). In an effort to link cultured C6 cells to molecular biological factors, demonstration of cell markers was attempted in these cells. MATERIALS AND METHODS: C6 cells were cultured in MEN solution and morphological changes were observed for 10 days. Next, cultured C6 cells were treated by temozolomide (TMZ), starting Day 3, and observed for 7 days. C6 cells were also implanted into rat's brain to observe tumor formation. CD133 and CD15 staining was done in cultured cells and TMZ treated cells. RESULTS: Small long cells with single nucleus grew to spindle cells on Day 1 to 2. Spindle cells increased rapidly, forming syncytiums on Day 3 to 4. As syncytiums multiplied, mitotic cells began to appear on Day 5 to 8. By Day 9-10 multiple small round cells were packed in groups, and then typical undifferentiated GBM cell pattern developed. During TMZ treatment, the number of C6 cells remarkably decreased forming necrotic areas, and only large spindle cells survived, resembling spongioblasts, in 4 days after the TMZ started. Implanted C6 cells survived, forming a mass, and contained spindle cells. CD133 and CD15 were used to stain cell membrane as markers on cultured and TMZ-treated cells, and results will be reported both in cultured C6 cells, and implanted tumors. CONCLUSION: The spindle cells of C6 cells played an important role for tumor growth and tumor invasion. Cell marker staining will allow for concluding relationship of spindle cells to stem cells (if stained with markers) or precursors, in comparison with radiated medulloblatoma cells stained with CD133.