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This study was undertaken to evaluate the integration of transplanted choroidal plexus epithelial cells with oxygen glucose deprived (OGD) organotypic spinal cord slices. Organotypic spinal cord slices were normally cultured for 14 days, then were OGD for 60 min. They were divided into transplantation group and non-transplantation group. The choroidal plexus epithelial cells, which were cultured for 6–7 days, were labeled by 1,1’-dioctadecyl-3,3,3’,3’-tetramethyl-indocarbocyanineperchlorate(CM-Dil), and were identified by transthyretin (TTR) in immunocytochemistry. They were adjusted to the density of 0.5–1×107/ml. 2µl cells suspension were transplanted into OGD spinal cord slices in transplantation group. Subsequently, the slices were normally cultured for 3 days (NT 3 d group), 7 days (NT 7 d group) and 14 days (NT 14 d group) respectively. Accordingly, in non-transplantation group, 2µl basal medium was dripped to the OGD organotypic spinal cord slices, then the slices were normally cultured for 3 days (N 3 d group), 7 days (N 7 d group) and 14 days (N 14 d group). After 3 days and 7 days of transplantation, the level of LDH in the culture medium was tested by Elisa, and the ratio of caspase 3 active/ caspase 3 was tested by western blotting. After 14 days of transplantation, the differentiations into neurons and astrocytes, and the synapses were identified by immunofluorescence histochemistry. At the same time, the ratios of cell differentiations and synapses in new system, and the changes of MAPK signaling pathway were tested by western blotting. Our result shows that the choroidal plexus epithelial cells were well labeled by CM-Dil and were immune-stained by TTR in immunocytochemistry. The transplantation cells could differentiate into neurons and astrocytes, and there were some synaptophysin positive vesicles between transplantation cells and OGD organotypic spinal cord slices. The level of LDH and the ratio of caspase 3 active/ caspase 3 in the NT 7 d group was significantly lower than which in the N 7 d group (P |
