An update on high-yield hepatocyte isolation methods and on the potential clinical use of isolated liver cells

Puviani, A.C.; Ottolenghi, C; Tassinari, B; Pazzi, P.; Morsiani, E

doi:10.1016/s1095-6433(98)10109-5

Cited by 45 publications

(27 citation statements)

References 53 publications

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“…Hepatocyte isolation was achieved by collagenase solution and perfusion. Percoll solution was used for further purification of the cells (Puviani et al, 1998). The cell damage was much lower, and a high cell yield was obtained compared to other methods.…”

Section: Discussionmentioning

confidence: 99%

“…This method is based on numerous isolation techniques, such as collagenase perfusion of the liver (Puviani et al, 1998;Guguen-Guillouzo, 2002;Li et al, 2005;Park and Song, 2006), permeation of the liver with chelator solutions (Battle and Stacey, 2001;Kravchenko et al, 2002), and enzymatic digestion or mechanical dissociation of the liver parenchyma (Puviani et al, 1998). When the cell yield is low or selective removal of the non-parenchymal cells is needed, Percoll (Puviani et al, 1998) or Ficoll (Guguen-Guillouzo, 2002 solutions may be used for the purification of cells. One of the fundamental principles of this procedure is based on the notion that Ca 2+ ions play a pivotal role in cellular adhesion.…”

Section: Introductionmentioning

confidence: 99%

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Isolation and identification of bovine primary hepatocytes

Jiang

Li²,

Liu³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. The liver is a unique organ that is endowed with a plethora of specialized functions. Most of its functional traits are controlled by hepatocytes. Primary hepatocytes have been used widely in in vitro models to understand the biological processes occurring in the liver. There are a number of methods used to separate hepatocytes, but the cell activity and purity are much lower in this condition. On the basis of previous research, in this study, the two-step collagenase perfusion technique was used for isolating hepatocytes. The key proteins of hepatocytes, cytokeratin-18 (CK-18) and albumin (ALB), were used to identify cells, and their contents were evaluated by immunohistochemistry and Western blotting. The results showed that the isolated hepatocytes comprised more than 96% of the corresponding protein volume stability. Therefore, this method was demonstrated to be reliable for identifying hepatocytes.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Isolation and identification of bovine primary hepatocytes

Jiang

Li²,

Liu³

et al. 2013

Genet. Mol. Res.

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show abstract

“…The isolation method is schematized in Figure 1. Because cell density and viability obtained by this simple enzymatic method are lower than with the perfusion method, Percoll solution was used for the further purification of the cells (Puviani et al 1998). It is possible to separate parenchymal cells from nonparenchymal and dead cells, and obtain pure hepatocyte populations by using Percoll.…”

Section: Discussionmentioning

confidence: 99%

“…In order to isolate hepatocytes from the liver, numerous isolation techniques such as collagenase perfusion of the liver (Puviani et al 1998;GuguenGuillouzo 2002;Li et al 2005;Park and Song 2006), permeation of the liver with chelator solutions (Battle and Stacey 2001;Kravchenko et al 2002), enzymatic digestion or mechanical dissociation of the liver parenchyma have been used in the literature (Puviani et al 1998). When the cell yield is low or selective removal of the non-parenchymal cells is needed, Percoll † (Puviani et al 1998) or Ficoll (GuguenGuillouzo 2002) solutions may be used for the purification of the cells.…”

Section: Introductionmentioning

confidence: 99%

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A cost-effective and simple culture method for primary hepatocytes

Aday

Hasırcı

Gürhan

2011

Animal Cells and Systems

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Hepatocytes, the major epithelial cells of the liver, maintain their morphology in culture dishes coated with extracellular matrix (ECM) components such as collagen and fibronectin or biodegradable polymers (e.g. chitosan, gelatin). In these coated dishes, survival of cells and maintaining of liver-specific functions may increase. The aim of this study was to determine a suitable, cost-effective and simple system for hepatocyte isolation and culture which may be useful for various applications such as in vitro toxicology studies, hepatocyte transplantation and bioartificial liver (BAL) systems. In order to obtain primary cultures, hepatocytes were isolated from liver by an enzymatic method and cultured on plates coated with collagen, chitosan or gelatin. Collagen, gelatin-sandwich and gelatin-cell mixture methods were also evaluated. Morphology and attachment of the cells were observed by inverted microscope and scanning electron microscope (SEM). An MTT assay was used to determine cell viability and mitochondrial activity.

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Liver Perfusion and Primary Hepatocytes for Studying Drug Metabolism and Metabolite Excretion

Xia

Balani

2011

Oral Bioavailability

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An update on high-yield hepatocyte isolation methods and on the potential clinical use of isolated liver cells

Cited by 45 publications

References 53 publications

Isolation and identification of bovine primary hepatocytes

Isolation and identification of bovine primary hepatocytes

A cost-effective and simple culture method for primary hepatocytes

Liver Perfusion and Primary Hepatocytes for Studying Drug Metabolism and Metabolite Excretion

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