[英语，研究]Matrix-based human hepatocyte transplantation

StephenW

<http://onlinelibrary.wiley.com/doi/10.1002/lt.22200/abstract>

Original Article

Primary human hepatocytes on biodegradable poly(l-lactic acid) matrices: A
promising model for improving transplantation efficiency with tissue
engineering†

Eva Török1,†, Marc Lutgehetmann2,3,†, Jeanette Bierwolf1, Stefan
Melbeck1, Jochen Düllmann4, Bjoern Nashan1, Peter X. Ma5,6,7, Joerg M.
Pollok1,*,‡Article first published online: 28 JAN 2011

DOI: 10.1002/lt.22200
Copyright © 2011 American Association for the Study of Liver Diseases
Issue

Liver Transplantation
Volume 17, Issue 2, pages 104–114, February 2011

Abstract
Liver transplantation is an established treatment for acute and chronic
liver disease. However, because of the shortage of donor organs, it does
not fulfill the needs of all patients. Hepatocyte transplantation is
promising as an alternative method for the treatment of end-stage liver
disease and as bridging therapy until liver transplantation. Our group has
been working on the optimization of matrix-based hepatocyte
transplantation. In order to increase cell survival after transplantation,
freshly isolated human hepatocytes were seeded onto biodegradable
poly(l-lactic acid) (PLLA) polymer scaffolds and were cultured in a flow
bioreactor. PLLA discs were seeded with human hepatocytes and exposed to a
recirculated medium flow for 6 days. Human hepatocytes formed spheroidal
aggregates with a liver-like morphology and active metabolic function.
Phase contrast microscopy showed increasing numbers of spheroids of
increasing diameter during the culture period. Hematoxylin and eosin
histology showed viable and intact hepatocytes inside the spheroids.
Immunohistochemistry confirmed sustained hepatocyte function and a
preserved hepatocyte-specific cytoskeleton. Albumin, alpha-1-antitrypsin,
and urea assays showed continued production during the culture period.
Northern blot analysis demonstrated increasing albumin signals. Scanning
electron micrographs showed hepatocyte spheroids with relatively smooth
undulating surfaces and numerous microvilli. Transmission electron
micrographs revealed intact hepatocytes and junctional complexes with
coated pits and vesicles inside the spheroids. Therefore, we conclude that
primary human hepatocytes, precultured in a flow bioreactor on a PLLA
scaffold, reorganize to form morphologically intact liver neotissue, and
this might offer an optimized method for hepatocyte transplantation because
of the expected reduction of the initial cell loss, the high regenerative
potential in vivo, and the preformed functional integrity.

Liver Transpl 17:104–114, 2011. © 2011 AASLD.

StephenW

原创文章

人肝细胞可生物降解聚（L -乳酸）矩阵：一个
希望提高移植效率的模型与组织
工程†

伊娃Török1，†，马克Lutgehetmann2，3，†，桃皮绒Bierwolf1，斯特凡
Melbeck1，约亨Düllmann4，Bjoern Nashan1，彼得十马5,6,7，约尔格米
Pollok1，*‡条第一发表时间：2011年1月28日

分类号：10.1002/lt.22200
版权所有© 2011年的美国肝病研究协会
发行

肝移植
第17卷，第2期，页104-114，2011年2月

摘要
肝移植是一个既定的治疗急性和慢性
肝病。但是，由于捐赠器官不足，它
不能满足所有病人的需要。肝细胞移植
作为一个有前途的终末期肝病治疗的替代方法
作为过渡，直到疾病和肝移植治疗。我们的集团
一直在研究矩阵为基础的肝细胞优化
移植。为了提高移植后的细胞存活，
新鲜分离的人类肝细胞接种于生物可降解
聚（L -乳酸）（聚乳酸）聚合物支架，并在流动培养
生物反应器。聚乳酸光盘与人类肝细胞接种和暴露于
6天中循环流动。人肝细胞形成球状
聚合体肝般的形态和积极的代谢功能。
相衬显微镜显示越来越多的球
增加在文化时期的直径。苏木精和伊红
组织学显示内的球体，完整可行的肝细胞。
免疫组化证实肝细胞功能和持续
保留肝细胞特异性细胞骨架。白蛋白，α- 1 -抗胰蛋白酶，
实验结果表明，尿素在养殖期间继续生产。
Northern杂交分析表明增加白蛋白信号。扫描
电子显微照片表明肝细胞球体相对平稳
表面起伏和无数的微绒毛。透射电镜
显微照片显示完整的肝细胞，并与交界物
小窝球体里面泡。因此，我们得出这样的结论
初级人类肝细胞，生物反应器预培养在流上的聚乳酸
脚手架，重组，形成完整的肝脏neotissue形态，并
这可能会提供一个优化的方法，因为肝细胞移植
最初的细胞损失预期减少，高再生
在体内的潜力，以及预制功能的完整性。

肝Transpl 17:104-114，2011。 © 2011年美国肝病学会。