Stem cells as a treatment for chronic liver disease and diabetes.
ABSTRACT Advances in stem cell biology and the discovery of pluripotent stem cells have made the prospect of cell therapy and tissue regeneration a clinical reality. Cell therapies hold great promise to repair, restore, replace or regenerate affected organs and may perform better than any pharmacological or mechanical device. There is an accumulating body of evidence supporting the contribution of adult stem cells, in particular those of bone marrow origin, to liver and pancreatic islet cell regeneration. In this review, we will focus on the cell therapy for the diseased liver and pancreas by adult haematopoietic stem cells, as well as their possible contribution and application to tissue regeneration. Furthermore, recent progress in the generation, culture and targeted differentiation of human haematopoietic stem cells to hepatic and pancreatic lineages will be discussed. We will also explore the possibility that stem cell technology may lead to the development of clinical modalities for human liver disease and diabetes.
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ABSTRACT: To rigorously test the in vivo cell fate specificity of bone marrow (BM) hematopoietic stem cells (HSCs), we generated chimeric animals by transplantation of a single green fluorescent protein (GFP)-marked HSC into lethally irradiated nontransgenic recipients. Single HSCs robustly reconstituted peripheral blood leukocytes in these animals, but did not contribute appreciably to nonhematopoietic tissues, including brain, kidney, gut, liver, and muscle. Similarly, in GFP+:GFP- parabiotic mice, we found substantial chimerism of hematopoietic but not nonhematopoietic cells. These data indicate that ``transdifferentiation'' of circulating HSCs and/or their progeny is an extremely rare event, if it occurs at all.Science 03/2003; 299(5611):1317; author reply 1317. · 31.20 Impact Factor
Article: Liver from bone marrow in humans.[show abstract] [hide abstract]
ABSTRACT: It has been shown in animal models that hepatocytes and cholangiocytes can derive from bone marrow cells. We have investigated whether such a process occurs in humans. Archival autopsy and biopsy liver specimens were obtained from 2 female recipients of therapeutic bone marrow transplantations with male donors and from 4 male recipients of orthotopic liver transplantations from female donors. Immunohistochemical staining with monoclonal antibody CAM5.2, specific for cytokeratins 8, 18, and 19, gave typical strong staining of hepatocytes, cholangiocytes, and ductular reactions in all tissues, to the exclusion of all nonepithelial cells. Slides were systematically photographed and then restained by fluorescence in situ hybridization (FISH) for X and Y chromosomes. Using morphologic criteria, field-by-field comparison of the fluorescent images with the prior photomicrographs, and persistence of the diaminiobenzidene (DAB) stain through the FISH protease digestion, Y-positive hepatocytes and cholangiocytes could be identified in male control liver tissue and in all study specimens. Cell counts were adjusted based on the number of Y-positive cells in the male control liver to correct for partial sampling of nuclei in the 3-micron thin tissue sections. Adjusted Y-positive hepatocyte and cholangiocyte engraftment ranged from 4% to 43% and from 4% to 38%, respectively, in study specimens, with the peak values being found in a case of fibrosing cholestatic recurrent hepatitis C in one of the liver transplant recipients. We therefore show that in humans, hepatocytes and cholangiocytes can be derived from extrahepatic circulating stem cells, probably of bone marrow origin, and such "transdifferentiation can replenish large numbers of hepatic parenchymal cells.Hepatology 08/2000; 32(1):11-6. · 12.00 Impact Factor
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ABSTRACT: Bone marrow stromal cells are progenitors of skeletal tissue components such as bone, cartilage, the hematopoiesis-supporting stroma, and adipocytes. In addition, they may be experimentally induced to undergo unorthodox differentiation, possibly forming neural and myogenic cells. As such, they represent an important paradigm of post-natal nonhematopoietic stem cells, and an easy source for potential therapeutic use. Along with an overview of the basics of their biology, we discuss here their potential nature as components of the vascular wall, and the prospects for their use in local and systemic transplantation and gene therapy.Stem Cells 04/2001; 19(3):180 - 192. · 7.70 Impact Factor
HEP (2007) 180:243–262
© Springer-Verlag Berlin Heidelberg 2007
N. Leviˇ car1(u) · I. Dimarakis1· C. Flores2· J. Tracey1· M. Y. Gordon2·
N. A. Habib1
1Department of Surgical Oncology and Technology, Faculty of Medicine, Imperial College
London, Hammersmith Hospital, Du Cane Road , London W12 0NN, UK
2Department of Haematology, Faculty of Medicine, Imperial College London,
Hammersmith Hospital, Du Cane Road, London W12 0NN, UK
Cell Therapy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Haematopoietic Stem Cells. . . . . . . . . . . . . . . . . . . . . . . . . .
Injured Liver . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.1Generation of Hepatocytes by Haematopoietic Stem Cells . . . . .
3.2Haematopoietic Stem Cell Therapy for Liver Diseases . . . . . . . .
3.2.1Animal Studies . . . . . . . . . . . . . . . . . . . . . . .
3.2.2Human Studies . . . . . . . . . . . . . . . . . . . . . . .
Diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4.1Haematopoietic Stem Cells forβ-Cell Generation . . . . . . . . . .
4.2Haematopoietic Stem Cells for Diabetes . . . . . . . . . . . . . . .
4.2.1Animal Studies . . . . . . . . . . . . . . . . . . . . . . .
4.2.2Human Studies . . . . . . . . . . . . . . . . . . . . . . .
Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Abstract Advances in stem cell biology and the discovery of pluripotent stem cells have
made the prospect of cell therapy and tissue regeneration a clinical reality. Cell therapies
better than any pharmacological or mechanical device. There is an accumulating body of
origin, to liver and pancreatic islet cell regeneration. In this review, we will focus on the
cell therapy for the diseased liver and pancreas by adult haematopoietic stem cells, as well
as their possible contribution and application to tissue regeneration. Furthermore, recent
progress in the generation, culture and targeted differentiation of human haematopoietic
stem cells to hepatic and pancreatic lineages will be discussed. We will also explore the
possibility that stem cell technology may lead to the development of clinical modalities for
human liver disease and diabetes.
Keywords Stem cell therapy · Haematopoietic stem cells · Liver disease · Diabetes
244N. Leviˇ car et al.
or enhance the biological function of damaged tissues or organs by utilising
cells and bioactive molecules to trigger, enhance, support and complement
the residual capacity for repair. This can be achieved by the transplantation of
cells, which are typically manipulated ex vivo, into a target organ in sufficient
numbers for them to survive long enough to restore the normal function of
organs and tissues. Possible candidate cells to be used include autologous
primary cells, cell lines, various stem cells including bone marrow stem cells,
cord blood stem cells and embryonic stem (ES) cells (Fodor 2003).
In recent years, advances in stem cell biology, including embryonic and
somatic stem cells, have made the prospect of tissue regeneration a potential
clinical reality, and several studies have shown the great promise that stem
cells hold for therapy (Assmus et al. 2002; Wollert et al. 2004). Despite the un-
questioned totipotency of ES cells, there are numerous unanswered biological
questions as to the regulation of their growth and differentiation. The safety
profile of unselected ES cells for transplantation early on demonstrated dys-
regulated cell growth resulting in teratoma formation (Reubinoff et al. 2000).
Moreover, the ethical and legal issues associated with ES cells have shifted
the focus to adult stem cells, and their regenerative potential has been under
The main role of adult stem cells, which are present in approximately
1%–2% of the total cell population within a specific tissue, is to replenish
of the tissue’s functional cells in appropriate proportions and numbers in re-
sponse to ‘wear and tear’ loss or direct organ damage (Fang et al. 2004). They
are vital in the maintenance of tissue homeostasis by continuously contribut-
ing to tissue regeneration and replacing cell lost during apoptosis or direct
injury (Li and Xie 2005). Adult stem and progenitor cells possess the capacity
of self-renewal and differentiation into one or more mature cell types. They
are able to maintain their populations within the human body through asym-
undifferentiated progeny (Preston et al. 2003). These properties make them
ideal candidates for stem cell-based therapies and tissue engineering.
Haematopoietic stem cells (HSC) are multipotent bone marrow cells that sus-
tain the formation of the blood and immune system throughout life. First
identifiedin 1961, HSC have beenby far the best characterisedand most stud-
Stem Cells as a Treatment for Chronic Liver Disease and Diabetes 245
ied example of adult stem cells (Till and McCulloch 1961). The bone marrow
stromal cells [called mesenchymal stem cells (MSC)] that have the ability to
2001). It was previously thought that adult stem cells were lineage restricted,
but recent studies demonstrated that bone marrow-derived progenitors in
addition to haematopoiesis also participate in regeneration of ischaemic my-
neurogenesis (Mezey et al. 2000).
into osteoblasts, adipocytes and chondrocytes (Pittenger et al. 1999).
Liver diseases impose a heavy burden on society and affect approximately
17% of the population. Cirrhosis, the end result of long-term liver damage,
has long been an important cause of death in UK and showed large rises
in death rates over the past 20 years (Ministry of Health 2001). The main
causes of cirrhosis globally are hepatitis B and C and alcohol abuse. Changing
patterns of alcohol consumption and the increasing incidence of obesity and
non-alcohol steatohepatitis (NASH) will continue to increase (Fallowfield and
Cirrhosis is a progressive liver disease and is marked by the gradual de-
struction of liver tissue over time. Persistent injuries lead to hepatic scarring
End-stage liver fibrosis is cirrhosis, whereby normal liver architecture is dis-
ruptedbyfibroticbands,parenchymal nodulesand vasculardistortion. Portal
hypertension and hepatocyte dysfunction are the end results and give rise to
plantation (OLT). However, the increasing shortage of donor organs restricts
liver transplantation. With the widening donor-recipient gap, the increasing
incidence of liver disease, life-long dependence on immunosuppression and
the poor outcome in patients not supported by liver transplantation, there is
obviously a demand for new strategies to supplement OLT.
of cells that can respond to liver injury and loss of hepatocytes (Sell 2001).
First, mature hepatocytes, which are numerous, respond to mild liver injury
by 1 to 2 cell cycles. Second, intra-organ ductal progenitor cells, which are
less numerous, respond by longer and limited proliferation. Third, stem cells
246N. Leviˇ car et al.
entering from the circulation participate in liver regeneration. These cells, in
injury (Petersen et al. 1999). In this latter mode, responding to severe injury,
they enter first as an intermediate cell population, which then mature into
New strategies for generating a viable source of healthy hepatocytes for
treating hepatic disorders are under investigation. Potential sources include
the expansion of existing hepatocytes, ES cells, progenitor/stem cells in the
liver, and bone marrow stem cells. Direct transplantation of hepatocytes has
already been successfully tried in a small number of patients as a bridge to
OLT or as a therapeuticalternative but there remain several limitations,which
mainly include limited hepatocyte amplification, the fact that replacement of
success fuelled the interest in haematopoietic stem cells for hepatic disorders.
In vitro studies have demonstrated the potential of bone marrow stem cells
to differentiate towards the hepatic lineage. Oh et al. (2000) and Miyazaki
et al. (2002) have shown that rat bone marrow contains a subpopulation (3%)
of cells co-expressing haematopoietic stem cell markers (CD34, c-kit, Thy-1),
α-fetoprotein (AFP) and c-met. They have also demonstrated the expression
of albumin, a marker of terminally differentiated hepatocytes, after culturing
crude bone marrow in the presence of hepatocyte growth factor (HGF) and
epidermal growth factor (EGF). Similar observations were made by Okumoto
et al. (2003), where rat bone marrow cells enriched for Sca-1 began expressing
that a subpopulation of murine mononuclear bone marrow cells isolated by
chemotaxis in response to theα-chemokine stromal-derived factor-1 (SDF-1)
expressed messenger RNA (mRNA) for AFP and a population enriched for
Sca-1 expressed mRNA for AFP, c-met and CK19. Purified murine HSC were
HNF1α) and mature hepatocyte markers (CK18, albumin, transferrin) when
are responsible for conversion (Jang et al. 2004).
into liver-like cells. When cultured on collagen matrix and in the presence of
Stem Cells as a Treatment for Chronic Liver Disease and Diabetes 247
Furthermore, when selected by SDF-1 chemotaxis they appear to be multi-
potent and express AFP (Kucia et al. 2004). Numerous cytokines and growth
commonly used in the majority of studies and have been shown to promote
hepatic differentiation in vitro (Block et al. 1996; Michalopoulos et al. 2003).
genitor cells (MAPC) has the potential to differentiate into hepatocyte-like
cells in the presence of HGF and fibroblast growth factor-4 (FGF-4) (Schwartz
et al. 2002). However, the MAPC culture is fastidious, with a substantial delay
between the isolation and the appearance of hepatocyte-like cells, which calls
into question their use in the clinic. In vitro hepatic differentiation of MSC in
the presence of HGF and oncostatin M was confirmed by another group (Lee
et al. 2004). Again, the differentiation process was lengthy but they demon-
strated the expression of liver-specific genes in differentiated cells and other
characteristics of liver cells, including albumin production, glycogen storage,
cytochrome P450 activity.
Liver has long been known to exhibit considerable regenerative potential,
but it has been only recently that we began to understand the implication of
stem/progenitor cells in this process. The extrahepatic stem cells such as HSC
are of particular interest since they are easily accessible.
Petersen et al. (1999) were first to show that bone marrow stem cells could
be a potential source of hepatic oval cells. The liver injury was induced with
hepatic cells were shown to be of bone marrow origin, demonstrated by using
markers for the Y chromosome, dipeptidyl peptidase IV enzyme, and L21-6
antigen to identify donor-derived cells.
The other demonstration of hepatocyte regeneration from bone marrow
et al. 1995), a lethal hereditary liver disease (Lagasse et al. 2000). Intravenous
and restored the biochemical function of its liver. The liver repopulation by
bone marrow cells was slow, although significant. The first hepatocytes of