本项目拟先用NT-3/TGF-α培养提纯、后用FACS技术分选人嗅粘膜OECs，同时直接分选嗅粘膜另外3种主要细胞（球状/水平基底细胞GBCs、HBCs和间充质干细胞MSCs），进而将GBCs、HBCs和OM-OECs分别与OM-OECs进行体外共培养，检测培养液中活性因子种类和含量并定量测定OM-OECs的NT-3基因表达以评价细胞活性，以揭示此3种细胞对OM-OECs活性的协同作用强度和机制，进而以人嗅球OECs做对照进行大鼠脊髓损伤体内验证，明确4种嗅粘膜来源细胞治疗SCI的修复作用强度。我们初步发现NT-3有助于OM-OECs的提纯，OM-MSCs与OM-OECs共培养具有协同促进作用，将用磁共振弥散张量成像技术对脊髓进行客观评价，并测其他细胞、生化和组织学指标。通过本研究可揭示人嗅粘膜GBCs、HBCs和MSCs在OECs治疗SCI中的协同作用机制，以提供移植用细胞新的来源选择。

Spinal cord injury (SCI) is one of the most difficult regeneration problems in the world. Cell transplantation is proved to be a promissing alternative for this condition. Olfactory ensheathing cells (OECs) transplantation has been demenstrated, in both basic and clinical research, to be safe and effective in SCI. However, up to now the OECs are mainly obtained from the allosome olfactory bulb (OB). Effect of cells from autologous olfactory mucosa (OM) on the treatment of SCI is attacting more and more attention now..This research focuses on four kinds of cells in OM namely Globose Basal Cells (GBCs), Horizontal Basal Cells (HBCs), Mesenchymal Stem Cells (MSCs) and Olfactory Ensheathing Cells (OECs). We firstly use Neurotrophin 3 (NT-3) or Transforming Growth Factor α (TGF-α) to culture and purify the OM-OECs from the OM mixed cells and then sort them out using the Flow cytometry (Fluorescence-activated cell sorting, FACS) according to the cell surface antigen (CSA) such as P75-NRT and GFAP. Because the other three kinds own stem cell properties, they are directly sorted out from the OM mixed cells according their own CSA (CD54 for HBCs, GBC-2 and BC-Ⅲ for GBCs and CD44 and CD105 for OM-MSCs, respectively). Furthermore, we will co-culture two kinds of cells, that is, one is OM-OECs and the other is GBCs, HBCs or OM-MSCs. So we could uncover the internal mechanism between the three kinds of cells and the OM-OECs and then go out the animal experiment. The animal experiments will be conducted using the spinal cord contusion model on the SD rats. These results will be compared with the OB-OECs. So we could confirm the different effects of the four kinds of cells on SCI. So far, we preliminarily found NT-3 could benefit the purification of OM-OECs which, however, need more efforts to confirm. And when co-cultrued, OM-MSCs could highlight the viability of OM-OECs extending longer protuberances while OM-OECs could activate the OM-MSCs differentiation to neural cells. Although we make hypothesis that they could affect each other through the cell direct contact or the active factors, the mechanism needs to be explore more. We will perform the Real-time PCR to quantitate the NT-3 gene expression to evaluate the OM-OECs' viability, which we have accumulate some experoence. And we will conduct Magnetic Resonance Diffusion Tensor Imaging (MR-DTI) to assess the spinal cord, which is very of sensitivity and accuracy in the living rats. Moreover, other measurments include cell couting, cell MTT, cell identification, serum diagnosis (inflammatory factor), active factors test (TGF-α, NT-3, BDNF, NGF and so on), spinal cord Oil dyeing and Western-blot (NogoA, Semaphorin3A, MAG, OMgP, NgR and RhoA). When the whole research finish we could clarify the effect and mechanism of GBCs/HBCs/OM-MSCs from human olfactory mucosa on OM-OECs transplantation treatment for SCI, which will offer a new approach to gain the seed cells for the transplantation treatment of SCI.