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《Journal of Chinese Medicinal Materials》 2007-12
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The Protective Effect of Astragalus membranaceus Extraction on Cryopreserved Primary-cultured Human Fetal Hepatocytes

WANG Yan1,ZHU Ying-qian2,LIN Jing-ming1,ZHANG Zhong-yi1,GAO Yi1(1.The Southern Medical University Affiliated Zhujiang Hospital,Guangzhou 510282,China;2.General Hospital of Tibet Military Command,Lasa 850003,China)  
Objective:To observe the influence of Astragalus membranaceus Extraction(AE) on the primary-cultured human fetal hepatocytes stored in liquid nitrogen and explore a new method for the cryopreservation of human hepatocytes with improved function.Methods:Human fetal hepatocytes were harvested by two-step collagenase perfusion,and then stored in a liquid nitrogen for one month with five different cryoprotectants(Ⅰ:10%DMSO,Ⅱ:5%DMSO+2mg/L AE,Ⅲ:5%DMSO+20mg/L AE,Ⅳ:5%DMSO+60mg/L AE,Ⅴ:5%DMSO+100mg/L AE).One month later,the cells were thawed rapidly and the viability,morphology and basic function of them were tested.Results:The human fetal hepatocytes in different groups showed various levels of viability,morphological manifestation and cell function respectively.After thawing,the viability rate and flash adhering rate in group Ⅳ and Ⅴ had no significant difference with group Ⅰ(P0.05),but were higher than group Ⅱ and Ⅲ(P0.05);the cell function analysis in the group Ⅳ,the results of ALB and AST level determination,NH4Cl transformation test,were the best among the groups(P0.05).Conclusion:AE can provide protection for human fetal hepatocytes in cryopreservation,and the best performance concentration level of its is 60mg/L;the preservation dosage of DMSO can be reduced when combined with AE in the preservation solution,which shows that AE has a synergistic effect with DMSO.
【Fund】: 广东省科技计划项目(2006B35802001)
【CateGory Index】: R285.5
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