Combined use of N-acetylcysteine and Liberase improves the viability and metabolic function of human hepatocytes isolated from human liver

Bartlett, David; Hodson, James; Bhogal, Ricky H.; Youster, Janine; Newsome, Philip N.

doi:10.1016/j.jcyt.2014.01.006

Cited by 16 publications

(11 citation statements)

References 35 publications

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“…Some studies have also found that NAC increased the level of glutathione in human hepatocytes, which may be responsible for the hepatocyte‐protective effects. Recently, NAC has been introduced for use in hepatocyte isolation . In the present experiment, administration of NAC in reperfusion largely ameliorated cell damage, as shown by reduced ALT and AST.…”

Section: Discussionsupporting

confidence: 50%

Comparison of methods for isolating primary hepatocytes from mini pigs

Wang

et al. 2016

Xenotransplantation

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Successful porcine hepatocyte isolation is crucial for the development of bioartificial liver devices and hepatocyte transplantation. Serva collagenase NB grades are formulated collagenases that are suitable for various tissue isolation applications. N-acetylcysteine (NAC) can improve the viability of human hepatocytes. The aim of this study was to compare the effectiveness of two collagenases and effect of NAC on hepatocyte isolation from porcine liver tissue. Porcine hepatocytes were isolated using the perfusion method from Bama mini pigs assigned to the Serva NB 4 group (n=6), the Serva NB 8 group (n=6), or the NB 8+NAC group (n=6). Viability and yield were defined as fresh hepatocytes and their spheroids formation after 24-hour rocker culture in serum-free medium. Metabolic function was assessed by gene expression, albumin, and urea synthesis. All procedures resulted in successful hepatocyte isolation. Cells from the NB 8+NAC group had (97.8±1.9)% viability, which was higher than the NB 8 group with (94.4±2.4)% and the NB 4 group with (94.5±3.2)% (P<.001). The final cell yield reached (11.8±1.0)×10(9) cells in the NB 8+NAC group, compared to (9.5±2.1)×10(9) cells in the NB 8 group (P<.01) and (9.1±1.1) ×10(9) cells in the NB 4 group (P<.001). The secretion of albumin was superior in the NB 8+NAC group at a concentration of (425.8±35.3) ng/mL compared to the NB 8 group (339.1±32.6) ng/mL (P <.001) and NB 4 group (293.6±43.3) ng/mL (P <.01). The injury of hepatocytes also decreased in the NB 8+NAC group (P<.01). The data are presented as means ± SD. Formulated collagenase Serva NB 8 and NAC could improve the porcine hepatocyte isolation, resulting in higher yields of viable cells.

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Section: Discussionsupporting

confidence: 50%

Comparison of methods for isolating primary hepatocytes from mini pigs

Wang

et al. 2016

Xenotransplantation

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“…With upgrade of commercial collagenase products such as Roche Liberase research grades and Serva collagenase NB grades, the isolations are more specific to the species, while the purity, viability, and yield of isolated primary hepatocytes are highly improved [4,18]. Except for the primary component, highly purified collagenaseand Ⅱmixture, we found that the amount of neutral protease thermolysin also determines the viability and cell yield, and especially the further success of spheroid formation.…”

Section: Hepatocyte Isolationmentioning

confidence: 87%

“…It also has direct antioxidant properties and appears to have hepatoprotective effects against liver ischemia/reperfusion injury to improve the viability [9]. It has been proved to improve the viability and conserves the metabolic function of hepatocytes [4,18].…”

Section: Hepatocyte Isolationmentioning

confidence: 99%

Isolations and Cultures of Primary Hepatocytes

Li¹,

Gao²,

Wu³

et al. 2017

J Clin Exp Pathol

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Various liver diseases result in liver failure, and liver transplantation and artificial liver support systems are emerging as alternative therapies for these disease. However, both treatments are depending on cell resources, so isolating sufficient number of functional primary hepatocytes is a most important early issue. Isolated hepatocytes are also a suitable system for the physiological, pharmacological and toxicological study of hepatic uptake, metabolism, excretion and toxicity. In this paper, the methods for hepatocyte isolation are reviewed; the culture and assessment methods are also examined. Hepatocyte transplantation and hepatocyte-based bioartificial liver support systems have attracted the attention of researchers. From a future perspective, developing of gene engineering is emerging as a promising way to modify cells, and this is renewing interest in the development of methods for isolation and culture of hepatocytes.

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“…Recent introduction of liberase, an enzyme used in pancreatic islet cell isolation, and N -acetylcysteine (NAC), an antioxidant with multiple hepatoprotective properties, into established isolation protocols has enabled improvements in overall isolation success from 40% to 70% (33, 50, 51). Additional strategies such as machine perfusion techniques and perfusate selection have been shown to dramatically increase cell yields (52, 53).…”

Section: Improving Human Hepatocytes Qualitymentioning

confidence: 99%

Clinical Hepatocyte Transplantation: What Is Next?

et al. 2017

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Purpose of review Significant recent scientific developments have occurred in the field of liver repopulation and regeneration. While techniques to facilitate liver repopulation with donor hepatocytes and different cell sources have been studied extensively in the laboratory, in recent years clinical hepatocyte transplantation (HT) and liver repopulation trials have demonstrated new disease indications and also immunological challenges that will require the incorporation of a fresh look and new experimental approaches. Recent findings Growth advantage and regenerative stimulus are necessary to allow donor hepatocytes to proliferate. Current research efforts focus on mechanisms of donor hepatocyte expansion in response to liver injury/preconditioning. Moreover, latest clinical evidence shows that important obstacles to HT include optimizing engraftment and limited duration of effectiveness, with hepatocytes being lost to immunological rejection. We will discuss alternatives for cellular rejection monitoring, as well as new modalities to follow cellular graft function and near-to-clinical cell sources. Summary HT partially corrects genetic disorders for a limited period of time and has been associated with reversal of ALF. The main identified obstacles that remain to make HT a curative approach include improving engraftment rates, and methods for monitoring cellular graft function and rejection. This review aims to discuss current state-of-the-art in clinical HT and provide insights into innovative approaches taken to overcome these obstacles.

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Combined use of N-acetylcysteine and Liberase improves the viability and metabolic function of human hepatocytes isolated from human liver

Cited by 16 publications

References 35 publications

Comparison of methods for isolating primary hepatocytes from mini pigs

Comparison of methods for isolating primary hepatocytes from mini pigs

Isolations and Cultures of Primary Hepatocytes

Clinical Hepatocyte Transplantation: What Is Next?

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