Cryopreservation and gel collagen culture of porcine hepatocytes

Liu, Hongling; Wang, Yingjie; Guo, Haitao; Wang, Yuming; Li, Jun; Yu, Yuecheng

doi:10.3748/wjg.v10.i7.1010

Cited by 4 publications

(1 citation statement)

References 45 publications

(45 reference statements)

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“…The most successful and well known of these products is University of Wisconsin (UW) solution. Naturally, these commercial solutions, as well as a handful of customized solutions, have been considered for cold storage of isolated hepatocytes, both in suspension (14,28,29) and in collagen gel culture (16,21). In contrast, the cold storage of hepatocyte spheroids is poorly studied (16).…”

Section: Introductionmentioning

confidence: 99%

Cold Storage of Rat Hepatocyte Spheroids

Liu

Glorioso

et al. 2014

Cell Transplant

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Cell-based therapies for liver disease rely on a high-quality supply of hepatocytes and a means for storage during transportation from site of isolation to site of usage. Unfortunately, frozen cryopreservation is associated with unacceptable loss of hepatocyte viability after thawing. The purpose of this study was to optimize conditions for cold storage of rat hepatocyte spheroids without freezing. Rat hepatocytes were isolated by a two-step perfusion method; hepatocyte spheroids were formed during 48 h of rocked culture in serum-free medium (SFM). Spheroids were then maintained in rocked culture at 37°C (control condition) or cold stored at 4°C for 24 or 48 h in six different cold storage solutions: SFM alone; SFM + 1 mM deferoxamine (Def); SFM + 1 mM cyclosporin A (CsA); SFM + 1 mM Def + 1 mM CsA, University of Wisconsin (UW) solution alone, UW + 1 mM Def. Performance metrics after cold storage included viability, gene expression, albumin production, and functional activity of cytochrome P450 enzymes and urea cycle proteins. We observed that cold-induced injury was reduced significantly by the addition of the iron chelator (Def) to both SFM and UW solution. Performance metrics (ammonia detoxification, albumin production) of rat hepatocyte spheroids stored in SFM + Def for 24 h were significantly increased from SFM alone and approached those in control conditions, while performance metrics after cold storage in SFM alone or cold storage for 48 h were both significantly reduced. A serum-free medium supplemented with Def allowed hepatocyte spheroids to tolerate 24 h of cold storage with less than 10% loss in viability and functionality. Further research is warranted to optimize a solution for extended cold storage of hepatocyte spheroids.

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Section: Introductionmentioning

confidence: 99%

Cold Storage of Rat Hepatocyte Spheroids

Liu

Glorioso

et al. 2014

Cell Transplant

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Differentiation-Promoting Medium Additives for Hepatocyte Cultivation and Cryopreservation

Gouliarmou

Pelkonen

Coecke

2014

Methods in Molecular Biology

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Isolated primary hepatocytes are considered as the reference system for in vitro hepatic methods. Following the isolation of primary hepatocytes from liver tissue, an unfavorable process named dedifferentiation is initiated leading to the attenuation of the hepatocellular phenotype both at the morphological and functional level. Freshly isolated hepatocytes can be used immediately or can be cryopreserved for future purposes. Currently, a number of antidedifferentiation strategies exist to extend the life span of isolated hepatocytes. The addition of differentiation-promoting compounds to the hepatocyte culture medium is the oldest and simplest antidedifferentiation approach applied. In the present chapter, the most commonly used medium additives for cultivation and cryopreservation of primary hepatocytes are reviewed.

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Sandwich-cultured hepatocytes: utility forin vitroexploration of hepatobiliary drug disposition and drug-induced hepatotoxicity

Bruyn¹,

Chatterjee²,

Fattah³

et al. 2013

Expert Opinion on Drug Metabolism & Toxicology

113

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When human (cryopreserved) hepatocytes are used to establish sandwich cultures, the model appears particularly valuable to quantitatively investigate clinically relevant mechanisms related to in vivo hepatobiliary drug disposition and hepatotoxicity. Nonetheless, the SCH model would largely benefit from better insight into the fundamental cell signaling mechanisms that are critical for long-term in vitro maintenance of the hepatocytic phenotype. Studies systematically exploring improved cell culture conditions (e.g., co-cultures or extracellular matrix modifications), as well as in vitro work identifying key transcription factors involved in hepatocyte differentiation are currently emerging.

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Cryopreservation and gel collagen culture of porcine hepatocytes

Cited by 4 publications

References 45 publications

Cold Storage of Rat Hepatocyte Spheroids

Cold Storage of Rat Hepatocyte Spheroids

Differentiation-Promoting Medium Additives for Hepatocyte Cultivation and Cryopreservation

Sandwich-cultured hepatocytes: utility forin vitroexploration of hepatobiliary drug disposition and drug-induced hepatotoxicity

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