Successful Isolation of Liver Progenitor Cells by Aldehyde Dehydrogenase Activity in Na"ve Mice

Abstract

The role of progenitor cells in liver repair and fibrosis has been extensively described, but their purification remains a challenge, hampering their characterization and use in regenerative medicine. To address this issue, we developed an easy and reproducible liver progenitor cell (LPC) isolation strategy based on aldehyde dehydrogenase (ALDH) activity, a common feature shared by many progenitor cells. We demonstrate that a subset of nonparenchymal mouse liver cells displays high levels of ALDH activity, allowing the isolation of these cells by fluorescence‐activated cell sorting. Immunocytochemistry and qPCR analyses on freshly isolated ALDH+ cells reveal an enrichment in cells expressing liver stem cell markers such as EpCAM, CK19, CD133, and Sox9. In culture, the ALDH+ population can give rise to functional hepatocyte‐like cells as illustrated by albumin and urea secretion and cytochrome P450 activity. ALDH1A1 expression can be detected in canals of Hering and bile duct epithelial cells and is increased on liver injury. Finally, we showed that the isolation and differentiation toward hepatocyte‐like cells of LPCs with high ALDH activity is also successfully applicable to human liver samples. Conclusion: High ALDH activity is a feature of LPCs that can be taken advantage of to isolate these cells from untreated mouse as well as human liver tissues. This novel protocol is practically relevant, because it provides an easy and nontoxic method to isolate liver stem cells from normal tissue for potential therapeutic purposes. (HEPATOLOGY 2012)

Podoplanin discriminates distinct stromal cell populations and a novel progenitor subset in the liver

“…This subset differences might reflect the phenotypic complexity and variance of ductular reaction observed in human liver diseases (14). In addition to this, the presented subset distribution is especially relevant, as previous studies mostly used a single surface marker to identify progenitor cells (6,8,9,16,27,38), barely reflective of the true composition or function of this cellular compartment. This is especially true for markers that are exclusively associated with one subset such as CD90, CD34, and Sca-1.…”

“…5A). Moreover, aldehyde dehydrogenase (Aldh1a1), an enzyme that activity enriched progenitor cells in healthy liver (6), and all tested genes involved in multiple pathways associated with stem/progenitor cell differentiation or maintenance such as members of the Notch, Wnt, hedgehog, and Hippo pathways (11,47) could be detected equally in both cell types in healthy liver (Fig. 5, A and B), indicating that both population belongs to the oval/progenitor compartment previously identified as CD133 ϩ cells in the liver (7,8,38).…”

Cancer Stem Cells and Aldehyde Dehydrogenase 1 in Liver Cancers

“…However, ALDH bright cells, including ALDH1 isoforms, evaluated using the Aldeluor assay, have been reported to be a marker of liver progenitor cells in the normal liver tissue [101] and of CSCs in HCC [102]. Interestingly, ALDH bright cells are atributed to ALDH1 activity.…”

Hepatic Progenitors of the Liver and Extra-Hepatic Tissues

“…The positive expression of aldehyde dehydrogenase (ALDH) has been used as a feature to select progenitors from adult mouse liver (53). ALDH + cells were shown to have stem cell characteristics and to express markers of human hepatic stem cells such as CD326, CK19, CD133, and Sox9.…”