Journal of Hepatology 49 (2008) 6–8

www.elsevier.com/locate/jhep

Editorial

A symphony of techniques for liver cell therapy,

only applicable to rats? q
Michael Ott
Clinic for Gastroenterology, Hepatology, Endocrinology and Twincore Center for Infectious Diseases,
Hannover Medical School, Feodor-Lynen Str. 7–9, 30625 Hannover, Germany

See Article, pages 99–106

The present issue of the Journal includes a study by
Kawashita et al., that investigates the potential of
immortalized hepatocytes to repopulate the liver of
Gunn rats after preparative irradiation and partial hep-
atectomy [1]. Primary hepatocytes from Gunn rats were
transduced with a thermolabile mutant simian virus 40
T-antigen as well as the human UGTA1a1 gene. The
transduced cells were transplanted into recipient Gunn
rats via intrasplenic injection. After various time points,
serum bilirubin concentrations, human UGT1A1 levels
and other parameters were measured. Three treatment
groups and one control group receiving the parental cell
line without transduction with the human UGTA1a1
gene were followed over a time period of 140 days.
The results demonstrate that the in vitro amplified and
transduced hepatocytes engraft, function and proliferate
in the recipient liver. Radiation therapy of the recipient
liver in combination with partial hepatectomy was most
effective in reducing serum bilirubin levels.
All single components of this study are well known
and have been published before. It was one of the many
achievements of Jayanta and Namita Roy-Chowdhury’s
laboratory to have thoroughly characterized the Gunn
rat as a model for cell and gene therapy [2,3]. Their lab-
oratory was also among the first to create and character-
ize immortalized hepatocytes, to study ex vivo gene
therapy for metabolic liver diseases and, in co-operation
Associate Editor: M. Trauner
q
The author declares that he does not have anything to disclose
regarding funding from industries or conflict of interest with respect to
this manuscript.
E-mail address: OTT-MHH@gmx.de
with other groups at the Albert Einstein College, to
apply preparative hepatic irradiation in combination
with hepatectomy to induce liver repopulation by trans-
planted cells. The present paper now orchestrates all the
pieces into one therapeutic approach and shows that a
bilirubin glucuronidation deficiency state can be cor-
rected in rats.
The authors provide evidence that immortalized
hepatocytes can be expanded in culture and trans-
planted into a recipient liver. This is both, a surprising
and a promising result, since primary adult hepatocytes
trypsinized from collagen coated cell culture plates
hardly engraft in a recipient liver after transplantation.
Of course, favourable conditions were created for the
cells. Preparative irradiation of the liver combined with
partial hepatectomy has been shown before to provide a
favourable environment for cell engraftment and repop-
ulation and probably was the key for the generation of
sufficient liver mass derived from UGT1A1 gene cor-
rected Gunn rat hepatocytes [4,5]. Additional help for
reducing the bilirubin levels after transplantation was
provided by transducing the immortalized hepatocytes
with a retrovirus expressing the UGT1A1 gene. Com-
pared to freshly isolated ‘‘wild-type” hepatocytes, the
transduced immortalized Gunn rat hepatocytes were
previously shown to over-express the UGT1A1 gene
by twofold [6]. In contrast, endogenously produced glu-
curonidation activity is likely to be down-regulated with
a gradual loss of the differentiated hepatic phenotype in
immortalized and expanded ‘‘wild-type” hepatocytes. It
would be interesting to know whether immortalized
‘‘wild-type” hepatocytes would be as efficient for the
correction of hyperbilirubinemia as isolated primary

0168-8278/$34.00 Ó 2008 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
doi:10.1016/j.jhep.2008.04.009
 M. Ott / Journal of Hepatology 49 (2008) 6–8
adult hepatocytes or gene corrected immortalized Gunn
rat-derived hepatocytes [7].
Will this strategy lead us to new horizons in the cell
therapy of metabolic liver diseases? Yes and no. One
of the major limitations for the development of liver cell
therapy programs is the shortage of human hepatocytes.
Many laboratories have focussed on stem cells as a
renewable source of hepatocytes, but overall, extensive
liver tissue formation has not been achieved with either
adult or embryonic stem cells. It is one of the main mes-
sages of this report that immortalized hepatocytes can
engraft, proliferate and correct a disease phenotype of
a recipient liver. Safety of the cells remains a concern.
Activation of the SV40 T-antigen is known to trigger
molecular events in the target cell, which are not fully
reversible by inactivation of the gene. Although tumours
have not been detected in the animals, activation of
alternative pathways inducing proliferation and new
technologies such as flanking transferred genes with
loxP targets for Cre recombinase mediated excision
may be needed for future clinical application [8].
It is obvious from the published data that hepatocyte
transplantation provides a therapeutic benefit in patients
with hereditary metabolic liver disease, but cannot com-
pletely and stably correct a disease phenotype [9,10].
Most patients, which are considered as targets for cell
therapy do not have signs of a liver pathology and thus
would not provide an intrinsic growth advantage for
transplanted cells. Although well established in surgical
departments, partial hepatectomy of the recipient liver
as part of a cell therapy protocol will probably not be
accepted for the often very young patients with meta-
bolic liver disease. Mild transient ischemia/reperfusion
damage of the liver may provide an alternative method
in order to increase engraftment and proliferation of
transplanted hepatocytes. In a conclusive study Malhi
and co-workers have demonstrated that 90 min of
ischemia and reperfusion in combination with prepara-
tive hepatic irradiation can result in massive repopula-
tion by transplanted cells [11]. The inherent risks of
the procedure may be significantly reduced by restriction
to single liver lobes. Cell transplantation combined with
regional ischemia/reperfusion of less than half of the
liver mass partially corrected hypercholesterolemia even
in the absence of radiation theraphy [12]. The radiation
dosage required for inhibition of hepatocyte prolifera-
tion would not necessarily impair liver function, but
would definitely cause DNA damage and carry the risk
for tumour formation. Stephenne et al. have recently
reviewed the application of radiation therapy in children
with liver organ transplantation and did not find an
increased risk for tumour formation or other patholo-
gies in the follow-up [13].
Would any of these procedures be too risky for
patients with hereditary metabolic liver disease? Lessons
may be learned from the hematologists. For years bone
7
marrow transplantation has been a common practice for
the treatment of congenital immune disorders and other
defects affecting bone marrow-derived cells. Complete
or partial myeloablation with chemotherapeutic drugs
is often part of the protocol to facilitate the engraftment
of donor bone marrow stem cells [14].
Advocacy for more aggressive cell therapy protocols
would have to consider that liver organ transplantation
is available for most of the children in the western world
and provides long-term correction of the metabolic
defect. Nevertheless, cell therapy concepts as an alterna-
tive or an additional approach to organ transplantation
may offer potential benefits for selected patients with
metabolic liver disease. It should be the objective of
the international scientific community to develop the
best protocols and techniques for the benefit of patients.
The fascinating experiments presented by Kawashita
et al. should help us to move closer to the routine appli-
cation of cell therapies for liver diseases.
References
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