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《Journal of Coloproctological Surgery》 2003-02
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The Isolation and Identification of Kupffer Cell in BALB/c Mouse

Zhang Sen. Coloproctological Surgery, the First Affiliated Hospital of Guangxi Medical University,Nanning, 530021 ;Wan Desen, Xiao Xibin. Tumor Hospital, Sun Yat sen University, Guangzhou,510060  
Objective:Liver is a vulnerable site of tumor metastases, but it contains abundant immunocyte(Kupffer Cell,KC) . This experiment probes to the isolation method of kupffer cell in BALB/c mouse in order to study its anti cancer activity.Methods:8 10 weeks health male BALB/c mice were selected, and with anaesthesia we got the liver and took away the connective tissue and blood.The liver was cut into pieces and 0.05%pronase E/0 04% EDTA were used alternatively. Filtration by stainless steel mesh was done before rinsing the cell and dissolving erythrocyte by Tris ammonia chloride. Then gradient centrifugation were done with 30% 70% Percoll and cultured in 10%FBS for 4 6 hours. The identification of kupffer cell was contrasted by the phagocytosis of ink and latex beads in vivo and in vitro.Results:The yield of kupffer cell is 7 57±1 92×10 6/mouse, namely 6 15×10 6/g Adhesive wall rate is 40 18%. Identification by 0 4% trypan blue shows that the cells survivel by more than 95% and the purity more than 90% in phagocytosis. The shape of kupffer cell is of multiplicity, irregularity, polygon and multiangular. Kupffer cell can survive for two weeks in vitro, but there is no phagocytic function at that time.Conclusion:It's a reliable means for isolating kupffer cell in BALB/c mouse by enzyme digestion process incorporation gradient centrifugation and adhibit wall culture, the work lays a foundation for study of liver metastasis.
【CateGory Index】: R329
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