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Title:: Improvement and simplification of method to culture neonatal SD rat cardiocytes

作者:: 王瑞华; 兰晓莉; 曹国祥; 安锐; 张永学; 华中科技大学同济医学院附属协和医院核医学科，湖北省分子影像重点实验室，湖北武汉 430022

Author(s):: WAGN Ruihua; LAN Xiaoli; CAO Guoxiang; AN Rui; ZHANG Yongxue; Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Hubei Key Laboratory of Molecule Imaging, Wuhan 430022, Hubei,China

关键词:: 新生SD大鼠; 心肌细胞; 细胞培养

Keywords:: neonatal rat; cadiocyte; cell culture

分类号:: R331;Q813.11

DOI:: -

文献标识码:: A

摘要:: 目的改良简化心肌细胞培养的条件和方法，培养纯度高、存活时间长的新生SD大鼠的心肌细胞。方法采用含牛血清白蛋白（bovine serum albumin，BSA）的Ⅱ型胶原酶单次消化心肌、差速贴壁1 h纯化心肌细胞进行培养，全过程在37℃ 条件下进行。于培养5、10、15、20、25、30 d时，用台盼蓝染色法检测细胞的存活率，并在生物倒置显微镜下观察形态学变化。取培养48 h的细胞，用免疫组织化学染色的方法检测心肌细胞的纯度。结果心肌组织在6 h时基本消化完全。培养5、10、15、20、25、30 d时，用台盼蓝染色法检测心肌细胞的存活率均大于95％，培养30 d时仍为95.2％。在生物倒置显微镜下观察，培养24 h左右的细胞之间形成交联并有细胞搏动。培养48 h的细胞经免疫组织化学染色法测定，心肌细胞的纯度为(97.1±1.9)％。结论应用改良并简化的心肌细胞培养方法和培养条件，可得到纯度和存活率均高能满足大多数科研要求的原代乳鼠心肌细胞。

Abstract:: AIM To culture purified neonatal rat cardiocytes using a simplified and modified method. METHODS Neonatal rat heart tissue was digested once in collagenase typeⅡincluding BSA and the purified cardiocytes were obtained by differential adhesion after 1 hour. All the steps were carried out in 37℃. The survival rate was assessed by typsin staining on days 5, 10, 15, 20, 25 and 30. The cell appearance was observed under inverted biological microscope and the purity of the cardiocytes was detected after 48 hours by immunohistochemistry. RESULTS The heart tissue was digested almost completely at 6 hours. The survival rate was more than 95% on days 5, 10, 15, 20, 25 and 30. Under the inverted biological microscope, we found that the cells connected to each other and beat after 24 hours. Immunohistochemistry showed that the purity of the cardiocytes cultured for 48 hours was (97.1±1.9)％. CONCLUSION By our simplified and modified method, we have successfully cultured purer neonatal rat cardiocytes.

参考文献/References

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新生SD大鼠心肌细胞培养方法的简化及改良(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

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