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比较80%和50%两种纯度的嗅鞘细胞在体外培养条件下的存活与生长,为嗅鞘细胞体内移植选用最佳纯度提供依据。分离、培养并纯化成年SD大鼠嗅鞘细胞,将纯化的嗅鞘细胞和收集到的贴壁成纤维细胞进行约80%和50%的比例混合后接种,继续培养1d或3d。所有细胞于不同时间点行P75免疫细胞化学荧光染色,以鉴定嗅鞘细胞的初始及其后培养过程中数量和纯度的变化。观察细胞状态,计数细胞总数和P75免疫阳性细胞数目,求得各组嗅鞘细胞数量和纯度并行统计学分析。结果显示:两种不同纯度嗅鞘细胞在培养1d时形态正常,3d时纯度为50%的嗅鞘细胞胞体依然饱满,突起更加纤长,数量和纯度亦无明显下降。而纯度为80%的嗅鞘细胞在培养3d时已出现胞体萎缩和突起变短,数量从1d时每一观察区域内的35±13显著下降为23±5(P=0.02),但纯度未发生明显变化。结果表明50%纯度嗅鞘细胞的存活及生长状态优于80%者,提示细胞体内移植时应考虑选取50%纯度的嗅鞘细胞。
The survival and growth of olfactory ensheathing cells in two kinds of purity of 80% and 50% were compared in vitro to provide the basis for selecting the best purity of the olfactory ensheathing cells for in vivo transplantation. Isolate, culture and purify adult SD rat olfactory ensheathing cells. The purified olfactory ensheathing cells and the collected adherent fibroblasts were inoculated at a ratio of about 80% and 50%, and cultured for 1 day or 3 days. All cells underwent P75 immunocytochemical staining at different time points to identify changes in the amount and purity of the olfactory ensheathing cells during the initial and subsequent culture. Observe the cell status, count the total number of cells and the number of P75 immunoreactive cells, calculate the number and purity of olfactory ensheathing cells in each group and analyze statistically. The results showed that the two kinds of olfactory ensheathing cells with different purity had normal morphology at 1 day of culture, and the cell body of olfactory ensheathing cells with 50% purity at 3d was still full with more slender protrusions and no obvious decrease in the number and purity. Olfactory ensheathing cells with a purity of 80% had atrophy and shortening of the cell body during the third day of culture, and the number decreased significantly from 35 ± 13 in each observation area to 23 ± 5 at 1 day (P = 0.02) Significant changes occurred. The results showed that the survival and growth of 50% pure olfactory ensheathing cells were better than 80%, suggesting that 50% purity of olfactory ensheathing cells should be considered when the cells were transplanted in vivo.