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目的：寻找一种简便、稳定、高产、经济的 Ｉｔｏ 细胞分离培养方法，为肝纤维化发生机制及抗肝纤维化药物的筛选提供一个可靠的细胞模型。方法：参照国内外介绍的实验方法并加以改良，采用体外两步灌注法及非连续密度分层液分离细胞。原代培养细胞，应用光镜、电镜、免疫组化和激发荧光实验观察鉴定。结果：细胞得率为３．３５×１０７／只，细胞存活率为９５％ ，纯度为９１％ 以上，优于国内外的报道。结论：本改良 Ｉｔｏ细胞分离培养方法简便、高产、经济、稳定，值得推广。
OBJECTIVE: To find a simple, stable, high yield and economical method for isolation and culture of Ito cells, and to provide a reliable cell model for the mechanism of liver fibrosis and the screening of anti-liver fibrosis drugs. Methods: Reference to the experimental methods introduced at home and abroad and to be improved, the use of two-step in vitro perfusion method and non-continuous density stratified separation of cells. Primary cultured cells, the application of light microscopy, electron microscopy, immunohistochemistry and excitation fluorescence experiments observed and identified. Results: The cell yield was 3.35 × 107 / cell, the cell survival rate was 95%, the purity was above 91%, which was better than the reports at home and abroad. Conclusion: The method of isolation and culture of this improved Ito cell is simple, high yielding, economical and stable and worth promoting.