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目的 建立可长时间维持细胞功能 ,减少细胞破坏的Kupffer细胞培养方法。方法 细胞破坏以乳酸脱氢酶(LDH)含量反映 ,细胞功能以一氧化氮 (NO)和胞内酸性磷酸酶 (ACP)水平反映。以双层夹心法 (doublecollagenculture ,DCC)对比普通培养法 (commonculture,CC)和单层胶原培养法 (singlecollagenculture ,SCC) ,在有或无血清状态下 ,Kupffer细胞功能维持和破坏情况。 结果 双层夹心法在培养 15d内 ,能降低培养上清LDH含量 ,维持NO和胞内ACP水平 ,并可在无血清状态下减少细胞破坏 ,维持细胞功能 ,与另两种方法相比作用显著。结论 双层夹心法对体外研究Kupffer细胞具有实际应用价值。
Objective To establish a Kupffer cell culture method that can maintain cell function for a long time and reduce cell destruction. Methods Cell destruction was characterized by lactate dehydrogenase (LDH) levels, and cellular function was reflected by levels of nitric oxide (NO) and intracellular acid phosphatase (ACP). The function of Kupffer cells was maintained and destroyed in the presence or absence of serum by comparing common culture (CC) and singlecollagen culture (SCC) with doublecollagenculture (DCC). Results The double-layered sandwich method could reduce the content of LDH in culture supernatant and maintain the level of NO and intracellular ACP within 15 days of culture, and could reduce cell destruction and maintain cell function in serum-free state with significant effect compared with the other two methods . Conclusion The double-sandwich method has practical value for the study of Kupffer cells in vitro.