乳鼠窦房结细胞原代培养与纯化方法的探讨

To Explore the Primary Pure-culture Method of Neonatal Rat's Sinoatrial Node Cells

目的:探讨乳鼠窦房结细胞的原代培养与纯化方法,观察细胞的生长状况,分析窦房结细胞的自律性及形态结构特征。方法:取SD乳鼠96只随机分为试验组和对照组,每组各48只,每次试验各取6只乳鼠窦房结组织,分别用前述两种方法进行细胞培养观察和比较两个培养组中各种细胞的数量及生长状况,以细胞的搏动频率评价细胞的活性,以梭形细胞的数量评价窦房结细胞的纯度。结果:(1)试验组明显减少了酶与细胞的接触时间和离心时间;简化了操作环节,优化了细胞的培养环境,可提高细胞的活性;(2)经过差速贴壁、阿糖胞苷处理,可明显减少不规则细胞的数量,窦房结细胞的数量明显上升(P<0.01)。结论:试验组纯化培养法是一种较为可行的乳鼠窦房结细胞原代培养方法,可为进一步研究提供高纯度、高活力的窦房结细胞。

Objective： To explore the primary pure-culture method of the neonatal rat＇s sinoatrial node（SAN） cells and to observe the vegetal status, beating frequence and morphological of the cells.Methods：96 Neonatal mouses was divided randomly into experiment group （differential velocity adherent technique and Cytarabine treated group） and contrasl group（common culture method group）. Observing the numbers and vegetal status of the cells in two groups. The activity of those cells was estimated through the beating freqence. The Purity of SAN cells was computed through the quantity of spindle cells. Results： （1） The experiment group distinctly reduced the contact time of enzyme and cells.Tbe centrifugal time has been shortened, too. It can significantly enhance the surviving frequence and vim through SimPlifing the experience processes and optimized the culture conditions of cells;（2） The proportion of sPindle cells increased obviously among cultured with differential attachment and cytarabine -treated method. It can reduce the ProPortion of Polygon ceils. Conclusion： The culture-method of experiment grouP is an ideal primary culture means of the sinoatrial node （SAN） cells, and it can supply higher Purified and viable SAN cells for further study.