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

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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, 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.