Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (12): 2161-2165.doi: 10.3969/j.issn.1673-8225.2011.12.018

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Culture of human hepatocyte L-02 at high concentration based on spherical porous chitosan microcarriers

Zhang Rui1, Han Bao-san2, Peng Cheng-hong2   

  1. 1Department of Oncosurgery, First Affiliated Hospital, Inner Mongolia Medical College, Hohhot  010059, Inner Mongolia Autonomous Region, China
    2Organ Transplantation Center, Rujin Hospital Affiliated to Medical College of Shanghai Jiao Tong University, Shanghai  200025, China
  • Received:2010-11-24 Revised:2011-01-06 Online:2011-03-19 Published:2011-03-19
  • About author:Zhang Rui, Doctor, Associate chief physician, Department of Oncosurgery, First Affiliated Hospital, Inner Mongolia Medical College, Hohhot 010059, Inner Mongolia Autonomous Region, China ruizhang999@sohu.com
  • Supported by:

    the National High-tech Research and Development Plan of China - the "863" Research Project, No. 2007AA02Z487*; the National Natural Science Foundation of China, No. 20074031*, 30672043* , 30772105*

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Abstract:

BACKGROUND: Recently, microcarrier culture as a high concentration cell culture technology has been used in hepatocyte culture in vitro.
OBJECTIVE: To periodically observe the morphological changes of human hepatocyte L-02 cultured by the spherical porous chitosan microcarrier.
METHODES: The self-made spherical porous chitosan microcarrier sample as a support to culture human hepatocyte L-02 as experimental group; non-spherical porous chitosan microcarrier as a support to culture human hepatocyte L-02 as control group. Cells in two groups underwent cell count, and the morphological changes were observed in experimental group, including inverted phase contrast biological microscope observation and scanning electron microscope observation.
RESULTS AND CONCLUSION: The quantity of the two groups’ cultured cells was increased in the former 3 days and reached the highest value at the third day. The cell quantity in experiment group was obviously higher than the number of cells without microcarrier culture in control group (P < 0.05). There was no significant difference of samples in experimental group (P > 0.05). The quantity of hepatocytes adhered to microcarrier surface was gradually increased in the former 3 days under inverted phase contrast biological microscope dynamic observation. There were lots of hepatocytes adhered to ball in the surface of a greater part of microcarrier, the total survive rate of hepatocyte was more than 90%, and hepatocyte kept a good morphological structure. Under scanning electron microscope, lots of hepatocytes adhered intensely each other in the surface, section, and inside of microcarrier. It is indicated that the self-made spherical porous chitosan microcarrier as a support in three-dimensional environment can undergo cell culture with high concentration.

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