Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (19): 3527-3532.doi: 10.3969/j.issn.1673-8225.2010.19.022

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Studies of Zuogui Wan medicated serum pharmacology based on the co-culture system of bone marrow stem cells and hepatocytes

Li Han-min, Gao Xiang, Yan Xue-sheng   

  1. Hepatopathy Institute, Affiliated Hospital of Hubei College of Traditional Chinese Medicine, Wuhan  430061, Hubei Province, China
  • Online:2010-05-07 Published:2010-05-07
  • About author:Li Han-min, Doctor, Chief physician, Doctoral supervisor, Hepatopathy Institute, Affiliated Hospital of Hubei College of Traditional Chinese Medicine, Wuhan 430061, Hubei Province, China lihanmin69@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 90709041*, 30672590*, 30271562*, 30371787*;
    the Key Basic Research Program of Science and Technology in China, No. 2002CCC00300*;
    the National Major Program in the 11th Five-Year Plan Period in China, No. 2008ZX10005-007*

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Abstract:

BACKGROUND: Previous studies have shown that Zuogui Wan medicated serum can promote transformation of bone marrow stem cells into hepatocytes. The confluence of bone marrow stem cells and hepatocytes can be eliminated, but interaction of bone marrow stem cells and hepatocytes in vivo cannot be simulated.

OBJECTIVE: To observe the effect of Zuogui Wan medicated serum on the transformation of bone marrow cells into hepatocytes.

METHODS: We established the co-culture system of bone marrow stem cells and hepatic tissue cells by using permeable supports (Transwell ® PTFE membrane insert) and established 10% Zuogui Wan medicated serum group and blank serum group. Medicated serum group added 10% Zuogui Wan medicated serum in the medium, whereas blank serum group added 10% normal rat serum in the medium. Bone marrow stem cells and hepatocytes were cocultured for 7, 14, 21, 28 and 35 days. Bone marrow cell slides in the permeable supports were gathered and glycogen was detected by PAS staining method. Hepatocyte markers (α-fetoprotein, CK18, albumin) were measured by immunocytochemistry method.

RESULTS AND CONCLUSION: Compared with blank serum group, the glycogen positive cell rate was obviously increased at the same time point (P < 0.01), and the α fetoprotein positive cell rate was significantly increased at 7 days after co-culture  (P < 0.01). At 14, 21, 28 and 35 days after co-culture, the α fetoprotein positive cell rate had a remarkable decrease (P < 0.05-0.01), and the positive cell rate of CK18 and albumin was apparently raised (P < 0.05). Using Zuogui Wan medicated serum in the co-culture system of bone marrow stem cells and hepatocytes was a better culture condition to promote bone marrow stem cell forming hepatocyte and maintain hepatocyte function.

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