中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (50): 9485-.doi: 10.3969/j.issn.1673-8225.2010.50.042

• 组织构建实验造模 • 上一篇    下一篇

乳兔窦房结定位、取材及纯化和培养

王庆志，周立，常玉巧   

  1. 新乡医学院人体解剖学教研室，河南省新乡市  453003
     
  • 出版日期:2010-12-10 发布日期:2010-12-10
  • 作者简介:王庆志☆，男，1970年生，河南省商城县人，汉族，2005年南方医科大学毕业，博士，副教授，主要从事心脏应用形态学研究。
  • 基金资助:

    河南省杰出青年科学基金(2005HANCET-15)；河南省卫生厅科研项目(200802008)。

     

Localization, cultivation and purification of sinoatrial nodes isolated from newborn rabbits
 

Wang Qing-zhi, Zhou Li, Chang Yu-qiao   

  1. Department of Anatomy, Xinxiang Medical University, Xinxiang  453003, Henan Province, China
  • Online:2010-12-10 Published:2010-12-10
  • About author:Wang Qing-zhi☆, Doctor, Associate professor, Department of Anatomy, Xinxiang Medical University, Xinxiang 453003, Henan Province, China wqzmgl@126.com
  • Supported by:

    the Science Fund for Distinguished Young Scholars of Henan Province, No. 2005HANCET-15*; the Science and Technology Research Program of Health Department of Henan Province, No. 200802008*

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摘要：

背景：分离和纯化培养的窦房结细胞是研究其超微结构及自律性的重要条件，然而，相应的培养方法尚未标准化。
目的：建立乳兔窦房结进行定位、取材和纯化培养方法，观察并判断培养窦房结细胞中起搏细胞的形态特点。
方法：新生24 h内乳兔心脏连续石蜡切片，苏木精-伊红染色，光镜下判断窦房结的位置并测定，计算其大小，光镜、电镜下观察纯化培养的窦房结细胞形态。
结果与结论：乳兔窦房结位于上腔静脉根部前壁向下至界沟后外约0.32 mm处。培养的窦房结细胞主要有3种形态细胞：梭形、蜘蛛形、多边形，其中梭形细胞最多，占(59.6± 7.3)%，搏动频率最快为(145±9)次/min，肌原纤维稀少，细胞器不发达。蜘蛛形和多边形细胞则和培养的心房肌细胞培养无差异。沿上腔静脉根部前壁向下至界沟后外侧可较精确地对窦房结进行取材。培养的乳兔窦房结细胞中，细胞体积小，搏动频率快，所占比率高的梭形细胞是窦房结的起搏细胞。

关键词: 细胞培养, 窦房结, 上腔静脉, 定位, 乳兔

Abstract:

BACKGROUND: The method of culture purified sinoatrial node cell is important in investigating its ultrastructural characteristics and autorhythmic mechanisms. However, the corresponding method has not been standardized.
OBJECTIVE: To summarize the localization, cultivation and purification of sinoatrial nodes isolated from newborn rabbits, and to study the morphological characters of primary cultured pacemaker cells.
METHODS: Hearts of the newborn rabbits (within 24 hours) were embedded in paraffin for hematoxylin-eosin staining. The location of sinoatrial nodes was observed under an optical microscope, the morphology of sinoatrial nodes cells were observed by light microscope and electron microscope.
RESULTS AND CONCLUSION: Sinoatrial nodes localized in the anterior wall of the superior vena cava and the posterior laterial wall of right atrium. There was about 0.32 mm between its lowest point and sulcus terminalis. Three distinct types of cells were observed among the cultured cells of sinoatrial nodes: spindle, spider and polygon. The spindle cells occupied the greatest proportion of the cultured cells (59.6±7.3)%. The spontaneous contraction frequency of spindle cells was the highest among the constracting cells (145±9) times per minute. The ultrastructure observation showed that myofibrils and other organelles in spindle cells were sparse and significantly decreased in number compared with triangle cells. There was no significant difference between triangle cells isolated from sinoatrial nodes and from atrial muscle. Sinoatrial nodes could be harvested along the anterior root of the superior vena cava down to the posterolateral sulcus. Among the cultured cells from neonatal rabbit sinoatrial nodes, the spindle cells with small body and fast pulse frequency are pacemaker cells.

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