Supernatant of rat C6 glioma cells induces human bone mesenchymal stem cells to differentiate into neuron-like cells / 中国组织工程研究

ABSTRACT

AIM: To investigate the differentiation of human mesenchymal stem cells (MSCs) into neuron-like cells induced by the supernatant of rat C6 glloma cells. METHODS: The heparin anticoagulant human bone marrow sampling was performed and human MSCs were separated flom human bone marrow using Percoll gradient method, followed by trypsinization and passage culture. When the MSCs at 4-6 passages reached a confluence of 90%, they were inoculated into a 24-well culture plate at a density of 2×103 per well. Cells were then divided into induction group and control group. In the induction group, C6 glioma cells were induced by the complete culture medium supplemented with 50% C6 glloma ceils supernatant (L-DMEM containing 0.1 fetal bovine serum), and half of the culture medium as replenished every 2 days. In the control group, cells were cultured in the complete culture medium. Both groups were treated 3 days and the expression of neuron-specific marker was monitored with S-P immunocytochemical stain. RESULTS: After an induction period of 24 hours, the differentiated MSCs showed typical neuron-like appearance, while the controlled cells exhibited no obvious alters. After 3 days of the induction, the neuron specific enola,se-positive rate of the cells in the induction was significantly higher than that in the control group (P < 0.01). The neurofilament protein-positive rate of cells in the induction group was (44.2±2.4)%, and the control group cells did not express neurofilament protein. Typical glial astrocyte marker glial fibrillary acidic protein was negatively expressed in both groups. CONCLUSION: Differentiation of MSCs into neuron-like cells can be induced by the supematant of rat C6 glioma cells in vitro.