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Title:: Experimental study of engineered cardiac patch in vitro

作者:: 常海霞; 郑强荪; 第四军医大学唐都医院心内科，陕西西安 710038

Author(s):: CHANG Hai-xia; ZHENG Qiang-sun; Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi′an，Shaanxi 710038, China

关键词:: 原代心肌细胞; 液态Ⅰ型胶原; 工程化心肌片层

Keywords:: primary cardiomyocyte; liquid collagen type I; engineered cardiac patch

分类号:: R540.27

DOI:: -

文献标识码:: A

摘要:: 目的探索在体外构建工程化心肌片层的方法。方法用胶原、Matrigel、2倍DMEM分别与制备的乳鼠原代心肌细胞混合，注入方形槽内形成心肌片层，5 d后行动态力学拉伸继续培养1周。常规HE染色、免疫组化染色等对组织工程化心肌组织，即心肌片层进行评价。结果心肌片层在培养1 d出现点状自发性跳动，随时间的延长跳动的区域逐渐增多,强度逐渐增大,3 d出现不一致的局部跳动，拉伸2 d出现肉眼可见整个片层的一致性的节律性跳动。HE染色显示片层中细胞排列整齐与正常成年大鼠心肌组织类似，免疫组化结果显示有丰富的心肌细胞。电镜显示条带中的心肌细胞含有丰富的肌原纤维,肌节结构和Z线清晰可见。结论该方法可以成功构建类似天然心肌组织的工程化心肌片层。

Abstract:: AIM To construct engineered cardiac patch in vitro. METHODS Liquid collagen type I, Matrigel and 2 DMEM were mixed with neonatal rat cardiac myocytes. The mixture was then pippetted into square molds to construct cardiac patch. Five days later, the cardiac patch was subjected to dynamic stretch for another 7 days. Microscopy and transmission electron microscopy, routine HE staining and immunohistochemical staining were used to analyze the EHTs. RESULTS Punctual spontaneous beating was seen in the cardiac patch at the first day of culture and more beating areas were seen thereafter. Areas with stronger and stronger beating and asynchronous beating were seen at the third day. The patch came to rhythmic synchronous beating at the second day after stretching. HE staining revealed that cardiac myocytes within the cardiac patch resembled those of the native adult cardiac tissue in morphology. Transmission electron microscopy revealed that the cardiac myocytes within the patch contained abundant myofibrils, clearly defined sarcomeres and Z lines. Histological and immunohistochemical analysis also showed more positive cardiac myocytes. CONCLUSION The method described in this study is helpful in constructing tissue engineered cardiac patch.

参考文献/References

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《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

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