The Preliminary Study On Isolation, Cultivation And Application Of Ofactory Ensheathing Cells

Spinal cord injury (SCI), a common disease in clinic rising in our country year after year, always happens among the adults. At present, there is no effective method to cure this disease. Most strategies for this disease mainly focus on how to prevent and reduce the secondly injury, and to help the adjustment of the recovery and regeneration of the central nervous system. The experiments show that together with gene therapy, the transplantation of embryo spinal cord, cells (stem cells) and the gene-transferred cells have achieved partial functional recovery of the injured spinal cord, which provide innovative methods for this disease. Especially, the olfactory ensheathing cells transplantation for treating the spinal cord injury has achieved preliminary success, which hopefully becomes one important method to resolve this century difficult question.This experiment mainly discussed the separation, cultivation, purification, proliferation and identification methods for the OECs from the mice, which also detect the roles of OECs in the growth of neurons. It aims at discovering rapid, stable and effective cultivation methods that will provide the cell source and theoretical instruction in the treatment of SCI by the OECs.We mainly applied four separation methods to isolate the OECs. All the cell suspension liquid was cultivated under the same condition. On the 12th day, we observed and compared the cells from the number and morphological aspect. After the purification of OECs through three different mentods, the purity of the cells was detected on the 12th day. We digested the OECs and the DRG (Dorsal root ganglion) and inoculated them in the same ratio (Density: 104/ml). Ten days later, under the invert microscope, we applied the morphological observation.It was demonstrated that after the mechanical cracking, trypsin digestion and following filtration, we obtained more OECs cells which were not damaged in the above separation process. The cultivated OECs always appeared as fusiform, process-bearing multipolar and round-flat (or fried egg-like) with significant morphological characters. The combination of differential velocity adherent and Ara-c inhibition included all their merits, which was proved as a simple and economical cultivation method for the OECs. OECs promoted the growth of DRG cells. Thus, the OECs can help the recovery of SCI, which needs further evidence.This experiment has discovered a mature protocol to separate and purify the OECs. It provides a stable cell resource to analyze the biological characters of OECs. Meanwhile, it also proved that OECs could promote the growth of other neurons, which provides a theoretical foundation for the treatment of SCI by the auto-transplantation of OECs.