Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (3): 473-477.doi: 10.3969/j.issn.1673-8225.2011.03.023

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Biological features of C3A hepatocytes versus L-02 immortalized hepatocytes under hypothermic stored condition

Li An-quan1, Li Qing-yong1,2, Zhang Qing-hua1, Jiang Zhi-xin1, Sha Hang1, Gao De-lu1, Gao Yi3   

  1. 1The 305 Hospital of Chinese PLA, Beijing   100017, China
    2Postgraduate College of Southern Medical University, Guangzhou   510515, Guangdong Province, China
    3Second Department of Hepatobiliary Surgery, Zhujiang Hospital Affiliated to Southern Medical University, Guangzhou   510253, Guangdong Province, China
  • Received:2010-10-25 Revised:2010-11-29 Online:2011-01-15 Published:2011-01-15
  • Contact: Zhang Qing-hua, Doctor, Professor, Doctoral supervisor, the 305 Hospital of Chinese PLA, Beijing 100017, China mdlqy@yahoo.com.cn
  • About author:Li An-quan, Associate chief physician, the 305 Hospital of Chinese PLA, Beijing 100017, China phdlqy@126.com
  • Supported by:

    the National High Technology Research and Development Program of China (863 Program), No.2006AA02A141*; the Special Fund of the Eleventh Five-year Plan for Military Medical Project, No. 08Z017*

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Abstract:

BACKGROUND: A large number of functional liver cells is the core of bioartificial liver. It is the existing research focus of bioartificial liver to explore a reliable method of cryopreservation of liver cells and to construct liver cell bank.
OBJECTIVE: To compare the biological characteristics of C3A hepatocytes that has entered clinical Ⅲ stage test and L-02 immortalized hepatocytes stored in UW solution at 4 ℃.
METHODS: The C3A hepatocytes and L-02 hepatocytes were adherent cultured, digested by 0.25% trypsinization, prepared into cell suspension, and stored in UW solution. At 0, 24, 48, 72 hours of hypothermic storage (4 ℃), the cell viability rate and cell apoptosis rate were measured using flow cytometry. Lactate dehydrogenase and aspartate aminotransferase release, the ability of hepatocytes to synthesize urea and secrete albumin were also determined.
RESULTS: The cell viability of C3A hepatocytes and L-02 hepatocytes was degraded following the time, but the cell viability of C3A hepatocytes was higher than L-02 hepatocytes (P < 0.01). Cell apoptosis was increased, but there was no difference between C3A hepatocytes and L-02 hepatocytes after 48 hours (P > 0.05). The lactate dehydrogenase and aspartate aminotransferase release were ascended, but the release of C3A hepatocytes was lower than L-02 hepatocytes (P < 0.01). The ability of hepatocytes to secrete albumin was decreased, but the ability of C3A hepatocytes was better than L-02 hepatocytes   (P < 0.01). The ability of hepatocytes to synthesize urea was also decreased, however, the ability of L-02 hepatocytes was better than C3A hepatocytes (P < 0.01). Results demonstrated that, the time of the C3A hepatocytes and L-02 hepatocytes which are hypothermicaly stored (4 ℃) in UW solution should not exceed 48 hours. The artificial liver prepared by C3A hepatocytes is more suitable for the liver function failure combined low albuminaemia, and artificial liver prepared by L-02 hepatocytes fits the liver function failure combined hepatic encephalopathy.

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