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背景:肝卵圆细胞具有强大的自我更新复制及在一定的条件下克隆增殖并分化为成熟肝细胞的能力,既可在体外用于构建生物人工肝的生物材料部分,也可以进行体内移植,还可为组织工程提供种子细胞,在治疗肝病方面具有广阔的前景。目的:建立成年Wistar大鼠肝卵圆细胞增殖模型,行体外分离和培养肝卵圆细胞,探索体外诱导其分化为肝细胞的可行性。设计:观察性实验。单位:南方医科大学南方医院消化病研究所。材料:实验于2003-12/2006-02在南方医院消化病研究所实验室完成。36只三四个月龄,其阳性率逐渐升高。④细胞化学方法显示诱导分化细胞胞浆G-6-P染色呈棕黑色沉淀,PAS染色呈红色颗粒。结论:乙硫氨酸灌喂联合2/3肝切除可复制成年Wistar大鼠的肝卵圆细胞增殖模型。胶原酶灌注结合Percoll密度梯度离心分离纯化可获得满意的肝卵圆细胞。大鼠肝卵圆细胞能在体外传代培养,在一定条件刺激下可诱导分化为肝细胞。
BACKGROUND: Hepatic oval cells possess strong self-renewal and replication ability to clone and proliferate and differentiate into mature hepatocytes under certain conditions. They can be used in vitro to build biomaterials of bioartificial liver, and can be transplanted in vivo, It also provides seed cells for tissue engineering and has a bright future in the treatment of liver diseases. OBJECTIVE: To establish a model of hepatic oval cell proliferation in adult Wistar rats, and to isolate and culture hepatic oval cells in vitro to explore the feasibility of inducing the differentiation into hepatocytes in vitro. Design: Observational experiments. Unit: Nanfang Hospital, Southern Medical University Institute of Digestive Diseases. MATERIALS: The experiment was performed in the laboratory of Institute of Digestive Disease, Nanfang Hospital from December 2003 to February 2006. 36 only three or four months old, the positive rate gradually increased. ④ Cytochemical staining showed that G-6-P staining in the cytoplasm of the differentiated cells was brownish-black and PAS was red granules. CONCLUSION: Ethionine can induce hepatic oval cell proliferation in adult Wistar rats with 2/3 hepatectomy. Collagenase perfusion combined with Percoll density gradient centrifugation to obtain satisfactory hepatic oval cells. Rat liver oval cells can be subcultured in vitro, under certain conditions can be induced to differentiate into hepatocytes.