Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (20): 3683-3687.doi: 10.3969/j.issn.1673-8225.2011.20.018

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Isolation, culture and identification of ventricular cardiomyocytes from embryonal, neonate and adult C57 mice

Zhang Ling1, Duan Ming-jun2, Wei Qin2, Chen Hua1, Shi Li3, Hou Yue-mei1   

  1. 1Arrhythmia VIP Laboratory, 2Section of Laboratory Animal Research, the First Affiliated Hospital of Xinjiang Medical University, Urumqi  830054, Xinjiang Uighur Autonomous Region, China; 3Department of Tumor, General Hospital of Chinese PLA, Beijing  100853, China
  • Received:2010-12-20 Revised:2011-02-19 Online:2011-05-14 Published:2011-05-14
  • Contact: Hou Yue-mei, Doctor, Professor, Doctoral supervisor, Arrhythmia VIP Laboratory, the first affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uyghur Autonomous Region, China houyuemei@sina.com Correspondence to: Shi Li, Department of Tumor, General Hospital of Chinese PLA, Beijing 100853, China
  • About author:Zhang Ling☆, Doctor, Arrhythmia VIP Laboratory, the first affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uighur Autonomous Region, China ydzhangling@126. com Duan Ming-jun, Section of Laboratory Animal Research, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uighur Autonomous Region, China Zhang Ling and Duan Ming-jun contributed equally to this paper.
  • Supported by:

    the High Technology Research and Development Program of Xinjiang Uighur Autonomous Region, No. 201010105*; the Open Subject of Key Laboratory of
    Angiocardiopathy of Xinjiang Uighur Autonomous Region, No. XJDX0903-2009- 03*

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Abstract:

BACKGROUND: Cultured cardiomyocytes are widely utilized in related researches such as physiological and toxicologic experiments, gene engineering, diseases model, drug screening and so on. How to harvest mouse cardiomyocytes with high activity is a key premise for these studies. 
OBJECTIVE: To establish a method to isolate and culture ventricular cardiomyocytes from embryonal, neonate and adult C57 mice.
METHODS: The ventricular myocardium from fetal, neonatal and adult C57 mice were minced and digested in trypsin, the purified ventricular cardiocytes were obtained by differential adhesion about 1 hour. The survival rate was assessed by trypan staining. Morphology of ventricular cardiomyocytes was observed through the methods of inverted phase contrast microscope, transmission electron microscope, scanning electron microscope, and the electrophysiological properties were identified through immunocytochemistry and microelectrode array.
RESULTS AND CONCLUSION: The ventricular tissues were almost completely digested through 3-6 times. Trypan staining showed that the survival rate of the cardiocytes cultured was more than 85%. Under the inverted phase contrast microscope, we found that the cells were fusiform or polyhedron and connected to each other and some cells began beat after 12 hours. The cells monolayer formed a network and beat spontaneously at the 30-90 times per minutes. The ventricular cardiomyocytes from embryonal, neonate and adult C57 mice can be obtained with well morphology and spontaneous beating with trypsin digestion.

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