两种含胶原凝胶的人工肝生物反应器的功能比较

徐兵; 吴林岚; 魏建威; 黄素钦; 陈怡; 赵芝萍; 蒋晓织; 郑登滋; 杨梅玉

doi:10.3969/j.issn.1001-5256.2014.10.013

两种含胶原凝胶的人工肝生物反应器的功能比较

DOI: 10.3969/j.issn.1001-5256.2014.10.013

1. 福建医科大学孟超肝胆医院肝病研究所
2. 福建省第二人民医院检验科

基金项目:

福州市科技局科研项目(2013-S-125-6)；

详细信息

中图分类号: R318.14
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出版历程
- 收稿日期: 2014-05-29
- 出版日期: 2014-10-20

Functional study on two artificial liver bioreactors with collagen gel

Institute of Liver Diseases, Mengchao Hepatobiliary Hospital, Fujian Medical University

Research funding:

摘要

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(70.1...
- 肝,人工 /
- 生物反应器 /
- 肝细胞 /
- 大鼠
Abstract:
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 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 SPSS13. 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 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) . Conclusion Hepatocytes suspended and immobilized in collagen gel might be more suitable for construction of a hollow fiber bioreactor of artificial liver.
- liver, artificial /
- bioreactor /
- hepatocytes /
- rats

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参考文献(13)

[1]YOU SL, LIU HL, RONG YH, et al.Preliminary investigation of hybrid bioartificial liver support system in treatment of HBV-related-acute-on-chronic liver failure[J].J Clin Hepatol, 2013, 29 (9) :685-688. (in Chinese) 游绍莉, 刘鸿凌, 荣义辉, 等.混合型生物人工肝治疗HBV相关慢加急性肝衰竭患者的初步探讨[J].临床肝胆病杂志, 2013, 29 (9) :685-688.

[2]YANG B, LIU BL, SHEN L.Comparison of two different in vitro primary human hepatocytes culture[J].Chin J Biological Eng, 2012, 31 (6) :941-945. (in Chinese) 杨波, 刘宝林, 沈力.两种体外人肝细胞的不同培养方式的比较研究[J].中国生物医学工程学报, 2012, 31 (6) :941-945.

[3]RAO XH, JIAN GD, ZHANG Z, et al.Biological metabolic function of the human liver cell line-1 used in bioartificial liver[J].Chin J Tissue Eng Res, 2013, 17 (18) :3295-3302. (in Chinese) 饶小惠, 简国登, 张志, 等.中国人肝细胞系1运用于生物人工肝的生物代谢功能[J].中国组织工程研究, 2013, 17 (18) :3295-3302.

[4]CHEN Z, WANG YJ, ZHANG SC, et al.Construction of polysulfone membrane bioreactor experimental system and its effects on plasma of patients with severe hepatitis[J].J Clin Rehabil Tissue Eng Res, 2008, 12 (6) :1175-1178. (in Chinese) .陈志, 王英杰, 张世昌, 等.聚砜膜生物反应器实验系统构建及对重型肝炎患者血浆的影响[J].中国组织工程研究与临床康复, 2008, 12 (6) :1175-1178.

[5]WANG YJ, LIU HL, WEN HW, et al.Collagen-gel immobilization culture of rat hepatocytes[J].Chin Hepatol, 2003, 8 (1) :15-17. (in Chinese) 王英杰, 刘鸿凌, 文红伟, 等.胶原凝胶固定培养大鼠肝细胞的功能与形态观察[J].肝脏, 2003, 8 (1) :15-17.

[6]XU B, JIANG JJ, PAN C, et al.Culture in vitro and abdominal transplantation of hepatocytes embedded in collagen gel[J].Chin J Organ Transplant, 2003, 24 (4) :229-230. (in Chinese) 徐兵, 江家骥, 潘晨, 等.胶原凝胶包埋肝细胞体外培养和腹腔移植[J].中华器官移植杂志, 2003, 24 (4) :229-230.

[7]ZHANG BF, ZHAO YS, GUO ZH, et al.In situ construction of injectable engineered liver tissue using collagen gel as scaffolds[J].J Clin Rehabil Tissue Eng Res, 2008, 12 (36) :7101-7104. (in Chinese) 张博峰, 赵云山, 郭振华, 等.以胶原凝胶为支架原位构建可注射型工程化肝[J].中国组织工程研究与临床康复, 2008, 12 (36) :7101-7104.

[8]LIU HL, WANG HF, WANG YJ, et al.Culture of cryopreservted hepatocytes of sucking pig with collagen gel[J].Med J Chin PLA, 2004, 29 (11) :967-970. (in Chinese) 刘鸿凌, 王慧芬, 王英杰, 等.冻存乳猪肝细胞的胶原凝胶培养[J].解放军医学杂志, 2004, 29 (11) :967-970.

[9]XU B, YANG XM, WU LL.Preliminary study on cryopreservation of mouse hepatocytes spheroids by ultra rapid vitrification[J].Chin Hepatol, 2012, 17 (2) :113-114. (in Chinese) 徐兵, 杨晓梅, 吴林岚.超速玻璃化法冻存小鼠肝细胞球形聚集体的初步研究[J].肝脏, 2012, 17 (2) :113-114.

[10] ZHANG J, DUAN ZP.Application of bioartificial liver systems in treatment of patients with liver failure[J].Chin J Hepatol, 2008, 16 (10) :728-730. (in Chinese) 张晶, 段钟平.生物人工肝在肝衰竭治疗中的应用[J].中华肝脏病杂志, 2008, 16 (10) :728-730. (in Chinese)

[11]LIU L, TANG SG.Establishment of an improved collagenase perfusion technique to isolate the hepatocyte effectively[J].J Clin Hepatol, 2012, 28 (3) :227-229. (in Chinese) 刘莉, 唐世刚.一种改良的胶原酶灌流高效分离肝细胞技术的建立[J].临床肝胆病杂志, 2012, 28 (3) :227-229.

[12]XU B, ZHAO ZP, QIU ZQ, et al.Establishment of the sterilization with ultraviolet and rolling and its application in sterilization of collagen solution[J].J Clin Rehabil Tissue Eng Res, 2009, 13 (25) :4886-4888. (in Chinese) 徐兵, 赵芝萍, 丘仲琼, 等.紫外线摇动灭菌法建立及在胶原溶液灭菌中的应用[J].中国组织工程研究与临床康复, 2009, 13 (25) :4886-4888.

[13]ZHAO J, LI XB, SU QH, et al.Three-dimensional culture of porcine hepatocytes for bioartificial liver[J].J Xi'an Med Univ, 2002, 23 (2) :136-140. (in Chinese) 赵军, 李晓斌, 苏清华, 等.用于生物人工肝的猪肝细胞三维立体培养[J].西安医科大学学报, 2002, 23 (2) :136-140.

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