Purification And Biological Activties Test Of Rats, Olfactory Ensheathing Cells

Objective1.Use modified differential attachment combine tryspin digestion method topurificate rats,olfactory ensheathing cells.2.Test the biological activities of olfactoryensheathing cells obtained from the above method.Methods Isolate olfactory ensheathing cells from7days old SD rats,olfactory bulbs,after grouped to purificate and culture cells by modified differential attachment(6h+18h)combine different time(1minute、3minutes、5minutes) tryspin digestion method, observemorphological characteristics and growth condition of olfactory ensheathing cells of differentgroups; Do immunohistochemical on low-affinity nerve growth factor receptor p75to assesscell purity; Use CCK-8to detect olfactory ensheathing cells,proliferation ability afterpurificated3days to15days; Use enzyme-linked immunosorbent to detect brain-derivedneurotrophic factor levels from different groups olfactory ensheathing cells after purificated3days to15days.Results Use modified differential attachment(6h+18h) combine different time(1min、3min、5min) tryspin digestion method to obtain olfactory ensheathing cells,purity are（57.52±2.13）%、（78.95±2.60）%、（49.01±0.74）%，pairwise comparisons between groupsshows P＜0.05,there is statistically significant; Olfactory ensheathing cells obtained afterpurificated7days to11days are all in the logarithmic growth phase, the results of cellnumber and brain-derived neurotrophic factor after purificated7days to11days comparisonsbetween groups shows3minutes group are the best, F=4.944and P＜0.05, there is statisticallysignificant. Conclusions1.Modified differential attachment(6h+18h) combine tryspin digestion3minutes can achieve higher purity olfactory ensheathing cells.2.Olfactory ensheathing cellsobtained after purificated7days to11days by this method are in the logarithmic phase, inwhich cell proliferation and brain-derived neurotrophic factor secretion ability are strongest.