OBJECTIVE: To investigate the influence of trauma on the tumorigenicity of rat glial tumor stem cells (C6-side population cell, C6-SP) in vivo. METHODS: Rat glial tumor stem cells C6-SP were isolated by flowcytometry. The biological behavior of C6-SP cells were observed by means of MTT experiment, single cell cloning, and cell cycle study with FCM CD133 expression was measured by immunofluorescence and FCM.. The tumorigenicity of C6-SP cells in vivo was evaluated by in situ tumor growth after intracranial implantation. The rat model was established by intracranial implantation of C6-SP cells. 10 days later, the rats in experimental groups were subjected to orthotopic or ectopic trauma. 24 days later, brain specimen was retrieved, gross tumor volume was measured, and Ki67 was evaluated by immunochemistry. The migration of stem cell was studied by the method to observe the relocation of C6-SP cells. RESULTS: Clustrus tumor growth was seen when C6-SP cell was cultured in serum-free medium. The doubling time of C6-SP cell was shorter than ordinary C6 cell. Single stem cell cloning efficiency of C6-SP cell was 77% whereas that of ordinary C6 cell was 16.4%. Among cloned C6-SP cell and ordinary C6 cell, 49.7% +/- 5.2% and 35.2 +/- 4.3% were at G0/G1 phase respectively. CD133 expression of C6-SP cells was positively shown by immunofluorescence. Tumorigenesis was 100% 2 weeks after in situ implantation of C6-SP cells. Gross tumor volume and Ki67 expression of orthotopic trauma group were larger and higher than those of ectopic trauma group or blank control group (P < 0.05) whereas difference was insignificant in the later two groups (P > 0.05). Red stained cells relocation was seen in both traumatized groups and absent in controls. CONCLUSION: C6-SP cells are the tumor stem cells (TSCs) for rat glioma. Trauma at the lesional site could increase tumorigenicity of the C6-SP cells. Moreover, trauma could induce migration of C6-SP cells in brain.
OBJECTIVE: To investigate the influence of trauma on the tumorigenicity of ratglial tumor stem cells (C6-side population cell, C6-SP) in vivo. METHODS:Ratglial tumor stem cells C6-SP were isolated by flowcytometry. The biological behavior of C6-SP cells were observed by means of MTT experiment, single cell cloning, and cell cycle study with FCMCD133 expression was measured by immunofluorescence and FCM.. The tumorigenicity of C6-SP cells in vivo was evaluated by in situ tumor growth after intracranial implantation. The rat model was established by intracranial implantation of C6-SP cells. 10 days later, the rats in experimental groups were subjected to orthotopic or ectopic trauma. 24 days later, brain specimen was retrieved, gross tumor volume was measured, and Ki67 was evaluated by immunochemistry. The migration of stem cell was studied by the method to observe the relocation of C6-SP cells. RESULTS:Clustrus tumor growth was seen when C6-SP cell was cultured in serum-free medium. The doubling time of C6-SP cell was shorter than ordinary C6 cell. Single stem cell cloning efficiency of C6-SP cell was 77% whereas that of ordinary C6 cell was 16.4%. Among cloned C6-SP cell and ordinary C6 cell, 49.7% +/- 5.2% and 35.2 +/- 4.3% were at G0/G1 phase respectively. CD133 expression of C6-SP cells was positively shown by immunofluorescence. Tumorigenesis was 100% 2 weeks after in situ implantation of C6-SP cells. Gross tumor volume and Ki67 expression of orthotopic trauma group were larger and higher than those of ectopic trauma group or blank control group (P < 0.05) whereas difference was insignificant in the later two groups (P > 0.05). Red stained cells relocation was seen in both traumatized groups and absent in controls. CONCLUSION:C6-SP cells are the tumor stem cells (TSCs) for ratglioma. Trauma at the lesional site could increase tumorigenicity of the C6-SP cells. Moreover, trauma could induce migration of C6-SP cells in brain.