The Phenotypic Characteristic of Liver-Derived Stem Cells From Adult Human Deceased Donor Liver

doi:10.1016/j.transproceed.2012.02.020

Transplantation Proceedings

Volume 44, Issue 4, May 2012, Pages 1110-1112

https://doi.org/10.1016/j.transproceed.2012.02.020 Get rights and content

Abstract

Liver transplantation is the only effective treatment for end-stage liver disease. Because of the limited donor availability, attention has been focused on the possibility to restore liver mass and function through cell transplantation. Stem cells are a promising source for liver repopulation after cell transplantation, but whether or not the adult liver contains hepatic stem cells is highly controversial. Several studies have suggested the presence of stem cells in the adult normal human liver. However, a population with stem cell properties has not yet been isolated. The purpose of this study was to identify and characterize progenitor cells in normal adult human liver. We isolated and expanded human liver stem cells (HLSCs) from a donated liver not suitable for liver transplantation or characterizing them by fluorescence-activated cell sorter, polymerase chain reaction, and immunofluorescence assay. HLSCs expressed the mesenchymal stem cell markers CD29, CD73, CD44, CD90, CD105, and CD166 but not the hematopoietic stem cell markers CD34, CD45, and CD117. HLSCs were also positive for vimentin and nestin, a stem cell marker. The absence of staining for cytokeratin-19, CD117, and CD34 indicated that HLSCs were not oval stem cells. In addition, HLSCs expressed CD26, and in a small percentage of cells, cytokeratin-8 and cytokeratin-18, indicating a partial commitment to hepatic cells. We concluded that HLSCs expressed several mesenchymal but not hematopoietic stem cell markers as well as CD26 and CK18, indicating a partial commitment to hepatic cells.

Section snippets

Isolation and Culture of HLSCs

This study was approved by our institutional review board, and all samples were obtained with informed consent. HLSCs were isolated from unused deceased donor whole liver that was not suitablet for liver transplantation. The donor was a 33 year-old female; the liver could not be transplanted because it was from a non–heart beating donor. Liver cell preparation was conducted using a slight modification of the two-step collagenase perfusion technique originally described by Seglen.⁴ In brief, the

Results

Morphologically, cultured HLSCs exhibited an elongated and spindled shape after 6 hours of seeding onto a culture flask (Fig 1). In an earlier study, fibroblastic features were described as a stem cell morphology.⁵ Characterization by flow cytometry revealed that the cells isolated by the described method were positive for stem cell markers CD29, CD73, CD44, CD90, CD105, and CD166, and negative for CD14 (monocyte), CD34 (heamatopoietic stem cell), and CD45 (leukocyte common antigen), indicating

Discussion

We identified in the adult human liver a cell population that fulfills the criteria for stem cell definition such as the capacity for self-renewal and multipotent differentiation. HLSCs expressed several mesenchymal but not hematopoietic stem cell markers and CD26 and CK18, indicating a partial commitment to hepatic cells. Isolation of a population of stem cells with similar biological potential from human liver tissue has been described earlier.2, 3, 6 HLSCs that were cultured contain distinct

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Am J Obstet Gynecol
(2006)
K.W. Lee et al.
Hepatocyte transplantation for glycogen storage disease type Ib
Cell Transplant
(2007)

There are more references available in the full text version of this article.

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Human liver tissue was obtained from a nontumor portion of the liver from a patient who received hepatic resection for hepatic hemangioendothelioma. Isolation of HLSCs was performed using a 2-step collagenase perfusion technique as previously described [3]. In the experiment, we used HLSCs from the 6th to 7th passages.
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Human liver tissue was obtained from a non-tumor part of liver from a patient who received hepatic resection for hepatic hemangioendothelioma. Isolation of HLSCs was performed using the two-step collagenase perfusion technique as previously described [12,13]. HLSCs at fifth passage were seeded into a fibronectin-coated 6-well plate at a density of 1 × 104 cells/cm2 and differentiated using the two-step protocol.
Liver transplantation is severely limited by donor shortage although it is the only effective treatment for end-stage liver disease. So the best alternative is hepatocyte transplantation. For obtaining human hepatocytes, some stem cells originating from extrahepatic or intraheptic tissues have been isolated and characterized. Previously we have reported that human liver-derived stem cells (HLSCs) could be isolated and expanded from donated livers unsuitable for transplantation; they expressed some markers of mesenchymal stem cells but neither hematopoietic nor oval cells. In this study, we isolated and expanded HLSCs with mesenchymal characteristics from another adult human liver. They showed mesenchymal morphology and grew well under serum condition similar to our previous reports. Also, they expressed some markers of mesenchymal stem cells, such as CD44, CD73, CD90, and CD105, through fluorescence-activated cell sorting analysis. When HLSCs were sequentially exposed to fibroblast growth factor−1 (FGF-1), FGF-4, and hepatocyte growth factor (HGF) followed by FGF-4, HGF, oncostatin M, and dexamethasone, they became round or polygonal, and expressed some hepatic markers such as albumin and α1-antitrypsin in the gene or protein level. Also, they showed urea synthesis activity 7 days after treatment of FGF-4, HGF, oncostatin M, and dexamethasone. These results provided that HLSCs would be a useful cell source in the field of regenerative medicine as well as liver cell biology.
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: Supported by a grant from the Korea Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (A080917).

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Experimental animal transplantationStem cells: LiverThe Phenotypic Characteristic of Liver-Derived Stem Cells From Adult Human Deceased Donor Liver

Abstract

Section snippets

Isolation and Culture of HLSCs

Results

Discussion

Exp Cell Res

Am J Obstet Gynecol

Hepatocyte transplantation for glycogen storage disease type Ib

Cell Transplant

Experimental animal transplantation
Stem cells: Liver
The Phenotypic Characteristic of Liver-Derived Stem Cells From Adult Human Deceased Donor Liver