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本文报道,通过局部灌流和振荡方法从外科肝活检标本分离成人肝细胞和进行原代培养的技术,用乙型肝炎病毒(HBV)接种培养的肝细胞,以证明 HBV 在此培养系中复制。从血清 HBsAg 和抗-HBc 阴性的肝胆疾病、肿瘤或外伤患者获得肝脏活榆标本,置于冰冷的培养液中,于37℃以含0.5mMEDTA 的无钙 Hanks 液用导管通过门静脉分枝灌注肝块5分钟。以此法去除血细胞后,再以0.05%胶原酶溶液(每升含 NaCl8.0g,KCl0.4g CaCl0.56g,NaH_2PO_40.78g,Na_2HPO_4·12H_2O 0.15g和 Hepes2.38g,试用前调至 pH7.5)子37℃灌注,流
Here we report the isolation of adult human hepatocytes from primary liver biopsy specimens by local perfusion and oscillatory techniques and the primary culture of hepatocytes. Hepatocytes were seeded with Hepatitis B virus (HBV) to demonstrate HBV replication in this culture. Liver live Elm specimens were obtained from serum HBsAg and anti-HBc-negative hepatobiliary diseases, tumors or traumatic patients and placed in ice-cold broth, perfusing the liver via a portal vein at 37 ° C with a calcium-free Hanks solution containing 0.5 mM EDTA Block for 5 minutes. After the blood cells were removed by this method, the cells were resuspended in 0.05% collagenase solution (containing 8.0 g of NaCl per liter, 0.56 g of KCl, 0.4 g of CaCl, 0.15 g of NaH 2 PO 4, 0.15 g of Na 2 HPO 4 · 12H 2 O and 23.8 g of Hepes, ) Sub 37 ° C perfusion, flow