味蕾细胞的Percoll梯度离心纯化技术研究

Development of a Discontinuous Percoll Gradient Centrifugation Process for Taste bud Cells Purification

以ICR小鼠的菌状味蕾细胞为研究对象,利用Percoll梯度离心技术纯化小鼠菌状味蕾细胞,通过台盼蓝染色和免疫组织化染色方法分别比较纯化前后的味蕾细胞存活率与纯度的变化,应用透射电镜从超微结构鉴定味蕾细胞。结果显示:通过不连续的Percoll密度梯度离心法对味蕾细胞进行纯化,细胞分层效果明显,主要位于体积分数20%与40%的Percoll工作液之间;免疫组化结果表明:纯化后味蕾细胞的纯度显著提高,从10%提高到65%(P<0.01),台盼蓝染色结果显示:纯化后细胞活度达94%,电镜则从超微结构上证实了纯化的细胞为味蕾细胞。

In this manuscript,the taste bud cells of fungiform papillae of ICR mice were select as research model and Percoll gradient centrifugation was used as purify method.The methods of trypan blue staining and immunohistochemistry staining were performed to compare the viability and purity of taste bud cells of before and after purification respectively.Taste bud cells were further identified by transmission electron microscopy.Results showed that taste bud cells,which were purified by discontinuous Percoll gradient centrifugation,were remarkably separated and mainly located interface between 20% and 40% Percoll working solution;the purity of taste bud cells were distinctly improved from less than 10% to more than 65%（p0.01） by this treatment;the viability of taste bud cells were up to 94% after purification by trypan blue staining and the results of transmission electron microscopy further confirmed taste bud cells from ultrastructure.