The Transplantation Of Olfactory Ensheathing Cells For Spinal Cord Injury And The Co-culture Of Olfactory Ensheathing Cells With Bone Marrow Mesenchymal Stem Cells In Vitro

Objective: There were three aims of the experiment: 1. To investigate the methods of isolation, culture, purification, proliferation and identification of olfactory ensheathing cells (OECs) from adult rats. 2. To investigate the possibility for the co-culture of olfactory ensheathing cells(OECs) with marrow mesenchymal stem cell(MSCs) in vitro and to study the biological characteristic in vitro. 3. To investigate the therapeutic effect of olfactory ensheathing cells on the spinal cord injury after transplantation.Methods: 1. The OECs were harvested from the olfactory bulb (OB) by three different isolation methods and were primary cultured in the same condition. After culturing 10 days, we made a contrast of cells quantity and cells morphology. 2.The OECs were purified by three different purification methods. On the tenth days, the cells purity was contrasted. 3. After isolation, culture and purification, the OECs were separated into two groups. One group was put into bFGF, the other group was not put in. every 2 days the cells quantity was counted, 10 days later, the cells quantity was compared. 4. The cells were identified by P₇₅ imunnofluorenscence.5.The OECs and MSCs were co-cultured with the same proportion after being digested, and the same density was 10⁴/ml. 10 days later, the morphological conformation of co-cultured cells was observed under phrase contrast microscope, the cells were identified by P₇₅ and BrdU imunnofluorenscence. But before identification the BrdU should be put into co-culture medium 2 days early. 6. The whole-section of T₁₀ spinal cord of adult S-D rats was made, and the rats were separated into three groups: the therapeutic group: OECs were transplanted into the spinal cord with microinjection methods after injury 14 days; the operation control group: the DMEM was injected with the same methods mentioned above; the normal group: the rats were not operated. 8 weeks later after transplantation, the weight of each group rats was observed by weighting every 2 weeks and the results were compared. The recovery of motor function was evaluated by modified Tarlov method and the results were compared. The pathological...