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两种含胶原凝胶的人工肝生物反应器的功能比较
Functional study on two artificial liver bioreactors with collagen gel
文章发布日期:2014年09月12日  来源:  作者:徐兵,吴林岚,魏建威,等  点击次数:1441次  下载次数:390次

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【摘要】:目的改进中空纤维型人工肝生物反应器。方法将大鼠肝细胞与胶原溶液混合并注入中空纤维反应器管外腔,待混合物在管外腔中形成胶原凝胶将肝细胞悬浮其中,构成胶原凝胶混悬肝细胞反应器(Ⅰ组);另将大鼠肝细胞混悬液注入涂有胶原层的中空纤维反应器管外腔,构成胶原涂层加肝细胞反应器(Ⅱ组)。两组反应器均由管内腔循环灌注培养液,置孵箱培养9 d,每24 h换液一次,每48 h取灌注液检测白蛋白(Alb)、尿素、乳酸脱氢酶(LDH)浓度,以检验反应器的肝细胞功能。采用统计软件SPSS 13.0进行统计学分析,计量数据以均数±标准差(x±s)表示,两组资料比较用配对t检验。结果Ⅰ组和Ⅱ组的Alb值均在灌注第3天达峰值,分别为 (1.41±0.08)和(0.65±0.05) g/L,3~9 d各时间点Alb值Ⅰ组均明显高于Ⅱ组(P值均<001)。Ⅰ组和Ⅱ组的尿素值分别在灌注第5天和3天达峰值,分别为(1.73±0.14)和(1.56±0.18)mmol/L,5~9 d各时间点尿素值Ⅰ组均明显高于Ⅱ组(P值均<0.01)。Ⅰ组和Ⅱ组的LDH值在灌注第9天达峰值,分别为(32.03±9.13)和(70.17±25.28)U/L,1~9 d各时间点LDH值Ⅰ组均明显低于Ⅱ组(P值均<0.01)。提示胶原凝胶混悬肝细胞反应器(Ⅰ组)比胶原涂层加肝细胞反应器(Ⅱ组)有更好的肝细胞合成代谢功能,肝细胞酶漏出也更少。结论胶原凝胶混悬固定肝细胞更适用于构建中空纤维型人工肝生物反应器。
【Abstract】:ObjectiveTo improve the hollow fiber bioreactor of artificial liver. MethodsRat hepatocytes mixed with collagen solution were injected into the external cavity of a hollow fiber reactor to construct a bioreactor of hepatocytes suspended in collagen gel (group Ⅰ). Other rat hepatocytes suspended in solution were injected into the external cavity of a hollow fiber reactor with a layer of collagen on the wall of the external cavity to construct a bioreactor of collagen layer and hepatocytes (group Ⅱ). For each group, the culture solution circulated through the internal cavity of the hollow fiber bioreactor; the bioreactor was put in a culture box for 9 d, and the culture solution in the internal cavity was exchanged for new one every 24 h; the concentrations of albumin (Alb), urea, and lactate dehydrogenase (LDH) in the culture solution samples were measured to examine the hepatocyte function of the bioreactor. Statistical analysis was performed using SPSS 130. Continuous data were expressed as mean±SD, and comparison between groups was made by paired t test. ResultsFor groups Ⅰ and Ⅱ, Alb levels reached peak values on day 3 of culture (1.41±0.08 g/L and 0.65±0.05 g/L); from day 3 to 9, group I had a significantly higher Alb level than group Ⅱ (t>7.572, P<0.01). For groups Ⅰ and Ⅱ, urea levels reached peak values on days 3 and 5 of culture (1.73±0.14 mmol/L and 1.56±0.18 mmol/L); from days 5 to 9, group I had a significantly higher urea level than group Ⅱ (t>8.418, P<0.01). For groups Ⅰ and Ⅱ, LDH levels reached peak values on day 9 of culture (32.03±9.13 U/L and 70.17±25.28 U/L); from days 1 to 9, group I had a significantly lower LDH level than group Ⅱ(t>5.633, P<0.01). Therefore, the bioreactor of hepatocytes suspended in collagen gel (group Ⅰ) showed a better hepatocyte function and less hepatic enzyme leakage compared with the bioreactor of collagen layer and hepatocytes (group Ⅱ). ConclusionHepatocytes suspended and immobilized in collagen gel might be more suitable for construction of a hollow fiber bioreactor of artificial liver.
【关键字】:肝,人工;生物反应器;肝细胞;大鼠
【Key words】:liver, artificial; bioreactor; hepatocytes; rats
【引证本文】:

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