Isolation and characterization of a novel population of progenitor cells from unmanipulated rat liver

Abstract

Widespread use of liver transplantation in the treatment of hepatic diseases is restricted by the limited availability of donated organs. One potential solution to this problem would be isolation and propagation of liver progenitor cells or stem cells. Here, we report on the isolation of a novel progenitor cell population from unmanipulated (that is, no prior exposure to chemicals and no injury) adult rat liver. Rat liver cells were cultured following a protocol developed in our laboratory to generate a unique progenitor cell population called liver-derived progenitor cells (LDPCs). LDPCs were analyzed by fluorescence-activated cell sorting, real-time polymerase chain reaction (RT-PCR), immunostaining and microarray gene expression. LDPCs were also differentiated into hepatocytes and biliary epithelium in vitro and examined for mature hepatic markers and urea and albumin production. These analyses showed that, LDPCs expressed stem cell markers such as cluster domain (CD)45, CD34, c-kit, and Thy 1, similar to hematopoietic stem cells, as well as endodermal/hepatic markers such as hepatocyte nuclear factor (HNF)3␤, hematopoietically-expressed homeobox gene-1, c-met, and transthyretin. LDPCs were negative for OV-6, cytokeratins (CKs), albumin, and HNF1␣. The microarray gene expression profile demonstrated that they showed some similarities to known liver progenitor/stem cells such as oval cells. In addition, LDPCs differentiated into functional hepatocytes in vitro as shown by albumin expression and urea production. In conclusion, LDPCs are a population of unique liver progenitors that can be generated from unmanipulated adult liver, which makes them potentially useful for clinical applications, especially for cell transplantation in the treatment of liver diseases. Liver Transpl 14: [333][334][335][336][337][338][339][340][341][342][343][344][345] 2008 Diseases of the liver are common causes of morbidity and mortality in the world. 1 Despite the high incidence of liver diseases that result in liver dysfunction and failure, current medical therapies are limited to supportive care, rather than curative approaches, with the possible exception of liver transplantation.Liver transplantation is considered to be the standard treatment for end-stage liver disease. 2 Unfortunately, its extensive application is restricted by the limited availability of donor organs. In addition, liver transplantation is associated with significant morbidity and mortality. As most liver disorders result from hepatocyte dysfunction, there has been great interest in transplantation of isolated hepatocytes. However, their clinical application is also dependent on the availability of good quality donor livers.To overcome the problem of limited donor organs and to make hepatocytes available for other applications, several approaches to isolate and propagate liver stem cells or progenitor cells have been developed. 3,4 It is

Potential hepatic stem cells reside in EpCAM+ cells of normal and injured mouse liver

“…However, it remained unclear whether such cells are present in normal adult liver. Cell lines with HSC characteristics have been established from normal mouse (Fougere-Deschatrette et al, 2006), human (Herrera et al, 2006) and rat (Sahin et al, 2008) liver. However, these cell lines were derived from a hepatocyte-enriched cell population by centrifugation using a Percoll gradient or from unfractionated cells, leaving their origin ambiguous.…”

Oxygenated Static Preservation of Donation after Cardiac Death Liver Grafts Improves Hepatocyte Viability and Function

“…Because most liver disorders result from hepatocyte dysfunction, HTx has attracted considerable attention [25,34]. HTx could alleviate the donor organ shortage if it was useful as a bridging therapy to OLTx [5,6,7,8].…”

“…The clinical application of the University of Wisconsin (UW) solution in the late 1980s considerably improved the quality and tolerance of grafts to cold ischemia [25]. Perfluorocarbon (PFC) liquids have been investigated as a medical application for many years.…”

“…These synthetic liquids are essentially bioinert and have no known deleterious histological, cellular, or biochemical effects [26,27,28,29,30]. The two-layer method (TLM) uses an oxygenated PFC and UW solution to preserve pancreases for whole organ and islet transplantation; TLM enhances pancreatic functions and increases islet cell yield after cold preservation compared with simple UW cold storage [25]. TLM directly oxygenates the graft by bubbling oxygen through PFC such that the graft continuously generates ATP, which is essential for cellular integrity [27,31].…”

Proliferation and differentiation potential of mouse adult hepatic progenitor cells cultured <italic>in vitro</italic>

“…But progress is hampered by difficulties in maintaining and expanding postnatal hepatic stem/progenitor cells and a lack of specific surface markers. Few studies reported successes in isolating hepatic progenitor cells from normal postnatal liver, which have been differently characterized by several markers, including CD34 [7,8], CD44 [9], c-kit [7,10], Thy-1 [7,11], CD133 [12,13], sca-1 [14], and side population cell attributes [15]. It suggests that these candidates of stem/progenitor cells in normal postnatal liver may represent different tissue-specific cell populations, because of various isolation protocols and different culture systems used.…”