不同血清浓度及培养时间对成年大鼠嗅黏膜嗅鞘细胞的影响

背景：课题组前期实验曾报道血清及神经生长因子对嗅鞘细胞体外增殖的影响，但对于嗅鞘细胞体外培养的血清浓度的选择及培养时间的掌握，这一方面的实验研究却报道较少 目的：通过研究不同血清浓度及体外培养时间对来源于成年大鼠嗅黏膜的嗅鞘细胞生长曲线的影响，从而了解不同实验条件对嗅鞘细胞体外生存和生长情况的影响。 方法：取成年大鼠嗅黏膜，进行体外分离培养，鉴定嗅鞘细胞，并用磺基罗丹明B酶标仪法测量各孔的吸光值（A值），转化为嗅黏膜嗅鞘细胞的生长曲线。 结果与结论：血清浓度0%组的A值明显低于其他浓度组。血清浓度10%-90%组细胞均可以正常生长。特别是血清浓度在10%-40%，对细胞生长起到明显的促进作用。随着细胞生长时间的延长，血清浓度0%组A值呈微弱的降低，其余各浓度组A值呈现增加状态。开始几天的A值增加趋势较弱，第9天之后增加趋势明显增强。血清浓度10%-40%组，在第13天，细胞生长迅速增快，第19天左右进入平台期，第23天A值有减弱的趋势。而血清浓度在50%-90%组，在第13天细胞生长达到峰值，随着时间的延长，A值逐渐变小。说明不同血清浓度及不同培养时间等体外培养环境对大鼠嗅黏膜嗅鞘细胞生长和生存有明显影响，因而利用血清进行体外细胞培养时应充分考虑所添加血清的浓度以及细胞培养的时间。

Influence of different blood serum concentration and growth time on olfactory ensheathing cells from the olfactory mucosa of adult rats

BACKGROUND：Preliminary experiments have reported the influence of serum and nerve growth factor on olfactory ensheathing cells proliferation in vitro, but there are less studies concerning choice of serum concentration and growth time for in vitro culture of olfactory ensheathing cells. OBJECTIVE：To find out the influence of different blood serum concentration and growth time on olfactory ensheathing cells from the olfactory mucosa of adult rats based on the growth curve of olfactory ensheathing cells. METHODS：The olfactory ensheathing cells from the olfactory mucosa of adult rats were separated, culture and identified in vitro. Sulforhodamine B and microplate reader were employed to measure absorbance values and plot growth curve of olfactory ensheathing cells. RESULTS AND CONCLUSION：When cultured for the same time in blood serum of different concentrations, absorbance values, especial y in the groups 10%, 20%, 30%, 40%, tended to increase with time except the 0%group. When cultured in the same serum for different time, absorbance values increased within the first 9 days, then promoted rapidly in the groups 10%, 20%, 30%, 40%at 13 days, entered the plateau phase at 19 days,＆nbsp;and decreased at 23 days;meanwhile, in the other groups （50%, 60%, 70%, 80%, 90%） the absorbance values peaked at the 13th day and then decreased gradual y. These findings indicate that different serum concentrations and different growth time in vitro affect cellgrowth and survival of olfactory ensheathing cells significantly, which should be ful y considered when cells are cultured in an in vitro condition.