CAJ | 학술논문

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。
목적：개진중공섬유형인공간생물반응기。방법장대서간세포여효원용액혼합병주입중공섬유반응기관외강，대혼합물재관외강중형성효원응효장간세포현부기중，구성효원응효혼현간세포반응기（Ⅰ조）；령장대서간세포혼현액주입도유효원층적중공섬유반응기관외강，구성효원도층가간세포반응기（Ⅱ조）。량조반응기균유관내강순배관주배양액，치부상배양9 d，매24 h 환액일차，매48 h 취관주액검측백단백（Alb）、뇨소、유산탈경매（LDH）농도，이검험반응기적간세포공능。채용통계연건SPSS 13．0진행통계학분석，계량수거이균수±표준차（x ±s）표시，량조자료비교용배대 t 검험。결과Ⅰ조화Ⅱ조적 Alb 치균재관주제3천체봉치，분별위（1．41±0．08）화（0．65±0．05）g/L，3～9 d 각시간점 Alb 치Ⅰ조균명현고우Ⅱ조（P 치균＜0.01）。Ⅰ조화Ⅱ조적뇨소치분별재관주제5천화3천체봉치，분별위（1．73±0．14）화（1．56±0．18）mmol /L，5～9 d 각시간점뇨소치Ⅰ조균명현고우Ⅱ조（P 치균＜0．01）。Ⅰ조화Ⅱ조적 LDH 치재관주제9천체봉치，분별위（32．03±9．13）화（70．17±25．28）U /L，1～9 d각시간점 LDH 치Ⅰ조균명현저우Ⅱ조（P 치균＜0．01）。제시효원응효혼현간세포반응기（Ⅰ조）비효원도층가간세포반응기（Ⅱ조）유경호적간세포합성대사공능，간세포매루출야경소。결론효원응효혼현고정간세포경괄용우구건중공섬유형인공간생물반응기。
Objective To improve the hollow fiber bioreactor of artificial liver.Methods Rat 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 inter-nal cavity was exchanged for new one every 24 h;the concentrations of albumin (Alb),urea,and lactate dehydrogenase (LDH)in the cul-ture 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.Results For groups Ⅰand 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 signif-icantly 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 hep-atocytes suspended in collagen gel (group Ⅰ)showed a better hepatocyte function and less hepatic enzyme leakage compared with the biore-actor of collagen layer and hepatocytes (group Ⅱ).Conclusion Hepatocytes suspended and immobilized in collagen gel might be more suitable for construction of a hollow fiber bioreactor of artificial liver.