来自细胞库的冷冻保存人肝细胞：体外功能及临床应用

Cryopreserved human hepatocytes from cell bank: in vitro function and clinical application.

作者信息

Baccarani U, Sanna A, Cariani A, Sainz M, Adani G L, Lorenzin D, Montanaro D, Scalamogna M, Piccolo G, Risaliti A, Bresadola F, Donini A

机构信息

Department of Surgery & Transplantation, University of Udine, Udine, Italy.

出版信息

Transplant Proc. 2005 Jan-Feb;37(1):256-9. doi: 10.1016/j.transproceed.2004.12.230.

DOI:10.1016/j.transproceed.2004.12.230

PMID:15808612

Abstract

UNLABELLED

We Aimed to analyze the in vitro function of isolated and cryopreserved human hepatocytes (CHH) from a cell bank and to define their potential clinical application in a bioartificial liver (BAL) device.

METHODS

Over 24 months, 103 not transplantable livers were utilized for human hepatocytes isolation and cryopreservation. Hepatocytes isolated by collagenase were analyzed for yield, viability, diazepam metabolism, and production of human albumin after isolation and cryopreservation in LN(2).

RESULTS

The causes for refusal for transplantation were macrosteatosis >60%, ischemic damage due to donor hypotension, and nonviral cirrhosis in 60%, 11%, and 8%, respectively. Cell yields averaged 7 million hepatocytes per gram of liver of mean viability of 80% +/- 13%. The viability of CHH after thawing averaged 50%. Thawed hepatocytes showed diazepam metabolism, and human albumin synthesis comparable to fresh cells. CHH were utilized as the biological component of a BAL for temporary support as three applications of two patients affected by fulminant hepatic failure awaiting urgent transplant. Ten to 13 billion viable CHH were loaded into each BAL. Liver function showed bilirubin and ammonia reduction at the end of each treatment. One patient was successfully bridged to emergency OLTx after one BAL; in the second case there was spontaneous recovery of liver function after two BAL.

CONCLUSIONS

Recovery of donor human livers unwanted for transplantation allowed isolation and cryopreservation of viable and functionally active human hepatocytes, which have been banked and successfully used for clinical applications of a BAL device.

摘要

未标注

我们旨在分析从细胞库中分离并冻存的人肝细胞（CHH）的体外功能，并确定其在生物人工肝（BAL）装置中的潜在临床应用。

方法

在24个月的时间里，103个不可用于移植的肝脏被用于人肝细胞的分离和冻存。对通过胶原酶分离的肝细胞在分离及在液氮中冻存后进行产量、活力、地西泮代谢及人白蛋白产生情况的分析。

结果

拒绝移植的原因分别为大脂肪变>60%、供体低血压导致的缺血损伤以及非病毒性肝硬化，分别占60%、11%和8%。细胞产量平均为每克肝脏700万个肝细胞，平均活力为80%±13%。冻融后CHH的活力平均为50%。冻融后的肝细胞显示出与新鲜细胞相当的地西泮代谢及人白蛋白合成能力。CHH被用作BAL的生物成分以提供临时支持，两名暴发性肝衰竭患者在等待紧急移植期间接受了三次BAL治疗。每次BAL中加载100至130亿个有活力的CHH。每次治疗结束时肝功能显示胆红素和氨水平降低。一名患者在接受一次BAL治疗后成功过渡到紧急肝移植；在第二个病例中，两次BAL治疗后肝功能自发恢复。