Experimental Study Of Venous Transplantation Of Olfactory Ensheathing Cells At Different Time Points To Repair Spinal Cord Injury

Objective 1:Labeling olfactory ensheathing cells with fluorescent QDs;2:To explore the optimal time window for transplantation of olfactory ensheathing cells for spinal cord injury.Methods 1:NASH differential attachment method was used to isolate and purify olfactory ensheathing cells derived from olfactory bulbs of SD rats.After successful extraction,the olfactory ensheathing cells were identified by immunofluorescence and the purity of olfactory ensheathing cells was determined.Different generations were detected by CCK-8 kit Cell growth and draw cell growth curve;2:The olfactory ensheathing cells were labeled with fluorescent quantum dots,and the toxicity of 10 nm,20nm,30 nm,40nm and other concentrations of quantum dot-labeled olfactory ensheathing cells was detected using the CCK-8 kit,and cell growth curves were plotted.Observation period marking efficiency;3:A hemisection model of spinal cord injury was established in the rat.A rat model was successfully selected.The olfactory ensheathing cells labeled with quantum dots were transplanted at 1D,3D,7D,and 10 D after spinal cord injury.At the spinal cord at the site,the amount of fluorescence collected at the lesion was detected by a small animal imager,and the olfactory ensheathing cells transferred to the lesion were detected by immunofluorescence.Results 1:The isolated olfactory ensheathing cells were extracted by differential adherence.Microscopic observation showed typical morphology of bipolar and tertiary olfactory ensheathing cells,and the cytoplasm had good light transmission.The calculated purity of olfactory ensheathing cells measured by immunofluorescence was greater than 85%;2:The CCk-8 method was used to detect the cell growth of different generations and draw the growth curve,and the P5 cell growth status was the best;3:After different concentrations of quantum dots are transfected into olfactory ensheathing cells,they have an inhibitory effect on cell growth.The greater the concentration of quantum dots,the stronger the inhibitory effect;4:Transfection of olfactory ensheathing cells with different densities with the same concentration of quantum dots has a high labeling rate,indicating that the size of the labeling rate has nothing to do with cell density;5:The olfactory ensheathing cells were transferred at different time points,and the small animal imager could detect the fluorescence accumulation at the injury site,and the fluorescence accumulation was the largest at 7D,which was statistically different from the other time point groups at P <0.05;6:The amount of olfactory ensheathing cells was measured by immunofluorescence method,and the number of cells in 7D was the largest.Conclusion 1:Fluorescent quantum dots can be used as tracers to label olfactory ensheathing cells;2:Olfactory ensheathing cells transplanted at different time points can be transferred to the spinal cord injury site;3:On the seventh day after spinal cord injury,the transplanted olfactory ensheathing cells have the most cells transferred to the injury site,which can be used as the best time window for transplanting olfactory ensheathing cells.