摘要
目的探讨用超速玻璃化法冻存的人肝组织（细胞）微团构建人工肝生物反应器的代谢和合成功能。方法将手术来源的人肝组织用胶原酶做有限消化，分解成肝组织微团，将肝组织微团用超速玻璃化法冻存，复苏后再用胶原酶有限消化成更小的肝细胞微团。将肝细胞微团与胶原溶液（pn7．4）混合，并将混合物注入中空纤维反应器管外腔，使混合物在管外腔形成胶原凝胶将肝细胞微团固定其中，构成生物反应器。用利多卡因循环灌注反应器管内腔24h，检验反应器的药物代谢功能；用乙型肝炎患者血浆循环灌注反应器管内腔6．5h，检测灌注液中血氨、间接胆红素（IBIL）、凝血酶原时间（PT）的变化，检验反应器的肝细胞功能。结果人肝组织微团经超速玻璃化冻存7d后复苏的肝细胞活率为（81．9±4．2）％。用利多卡因灌注24h后，利多卡因浓度为6．2mg/L，代谢转换率为92％。100mL患者血浆循环灌注生物反应器6．5h后，灌注液中血氨、IBIL和PT值分别为（22．5±6．1）μmol／L、（29.3±8．6）μmol／L和（20．5±2．7）s，低于灌注0．5h的相应值，差异均有统计学意义（t=13．5、17．9和7．4，P均〈0．05）。结论超速玻璃化法冻存的人肝组织（细胞）微团复苏后固定于胶原凝胶中构建的中空纤维型人工肝生物反应器，有较好的肝细胞代谢和合成功能。
Objective To develop an artificial liver bioreactor with collagen gel and the cryopreserved liver tissue micro- lumps （hepatocyte lumps）, and study function of metabolism and synthesis by ultra rapid vitrification. Methods Human liver tissues obtained from surgical operations were decomposed into micro-lumps by the limited digestion of collagenase. The micro-lumps were cryopreserved by ultra rapid vitrification and then revived and decomposed into smaller micro-lumps （hepatocyte lumps） by the limited digestion with collagensase. The hepatocyte lumps were mixed with collagen solution （pH 7.4） and injected into the external cavity of a hollow fiber reactor. The mixture would become a collagen gel in the reactor. Lidocaine and plasma of hepatitis patients were circulated through the inner cavity of the hollow fiber reactor for 24 h and 6.5 h, respectively to examine the function of the hepatocytic reactor （lidocaine concentration after circulation, blood ammonia, indirect bilirubin, and prothrombin time）. Results The viability of the hepatocytes of the cryopreserved liver tissue micro-lumps was （81.9~4.2）%. The concentration of lidocaine after 24-h circulation was 6.2 mg/L, with the metabolic coylversion of 92%. The values of blood ammonia, indirect bilirubin, and prothrombin time of the samples of 6.5-h circulation were （22.5±6.1） μmol/L,（29.3±8.6） μmol/L and （20.5±2.7） s, which significantly lower than those of 0.5-h circulation （t=13.5, 17.9 and 7.4, P all〈0.05）. Conclusions A hollow fiber reactor of artificial liver can be made from cryopreserved human liver tissue micro-lumps （hepatocyte lumps） by ultra rapid vitrification with collagen gel, and this reactor preserves good ability of hepatocytic metabolism and synthesis.
出处
《国际流行病学传染病学杂志》
CAS
2014年第5期295-299,共5页
International Journal of Epidemiology and Infectious Disease
基金
福州市科技局科研项目（2013-S-125-6）
关键词
肝
人工
生物反应器
肝组织
冻存
Liver, artificial
Bioreactor
Liver tissue
Cryopreservation
