人胚胎窦房结细胞原代培养及纯化方法探讨

Primary culture and purification method of human embryonic sinoatrial node cells

目的:探讨人胚胎窦房结细胞的原代培养及纯化方法,寻找一种较为可行的适宜于人胚胎窦房结细胞生长的方法。方法:取人工水囊引产的12～16周人胚胎心脏,随机分为实验组及对照组,取其窦房结组织分别进行细胞培养,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:实验组简化了操作环节,优化了细胞的培养环境,可提高细胞的活性;经过差速贴壁阿糖胞苷纯化处理,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<0.05)。结论:纯化培养法是一种较为可行的人胚胎窦房结细胞原代培养方法,可为进一步研究提供生长较稳定的窦房结细胞。

Objective：To explore the primary pure-culture method of human embryonic sinoatrial node（SAN） cells and observe the vegetal status,beating frequence and mor phological of the cells. Method： Take the artificial water sac induced labor human embryonic heart from 12 to 16 weeks,and divided into experiment group and control group.We observed 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. Result：①The experiment group distinctly reduced the contact time of enzyme and cells.The centrifugal time has been shortened too.It can signifcantly enhance the surviving rate through simplifying the experiemeny processes and optimized the culture conditions of cells;②The proportion of spindle cells increased obviously when cultured with differential attachment and cytarabine-treated method.It can reduce the proportion of polygon cells. Conclusion：The culture-method of experiment group is an ideal primary culture means of SAN cells,and it can supply higher stable and viable SAN cells for further study.