Effects of plasma from patients with fulminant hepatic failure on function of primary rat hepatocytes in three‐dimensional culture

Nagaki, Masahito; Naito, Tomoo; Ohnishi, Hiroo; Akaike, Toshihiro; Muto, Yasutoshi; Moriwaki, Hisataka

doi:10.1111/j.1478-3231.2005.01127.x

Cited by 3 publications

(3 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The 3D configuration and/or medium perfusion contribute in a large part to the increased plasma-resistance observed in BALs compared to monolayer cultures. This is in accordance with a previous report that primary rat hepatocytes are protected from plasma-induced toxicity by 3D-culturing [27]. The applied 3D culture platform provides a flexible culture surface, which may be essential for maintaining cell-cell contact.…”

Section: Discussionsupporting

confidence: 91%

HepaRG-Progenitor Cell Derived Hepatocytes Cultured in Bioartificial Livers Are Protected from Healthy- and Acute Liver Failure-Plasma Induced Toxicity

Wenum¹,

Treskes²,

Adam³

et al. 2018

Cell Physiol Biochem

View full text Add to dashboard Cite

Background/Aims: For applicability of cell-based therapies aimed at the treatment of liver failure, such as bioartificial livers (BALs) and hepatocyte transplantation, it is essential that the applied hepatocytes tolerate exposure to the patient plasma. However, plasma from both healthy donors and acute liver failure (ALF) patients is detrimental to hepatocytes and hepatic cell lines, such as HepaRG. We aimed to elucidate the underlying mechanisms of plasma-induced toxicity against HepaRG cells in order to ultimately develop methods to reduce this toxicity and render HepaRG-BAL treatment more effective. Methods: Differentiated HepaRG cells cultured in monolayers and laboratory-scale BALs were exposed to culture medium, healthy human plasma, healthy porcine plasma and ALF porcine plasma. Healthy human plasma was fractionated based on size- and polarity, albumin depleted and heat treated to characterize the toxic fraction. The cells were assessed for viability by total protein content and trypan blue staining. Their hepatic differentiation was assessed on transcript level through qRT-PCR and microarray analysis, and on functional level for Cytochrome P450 3A4 activity and ammonia elimination. Mitochondrial damage was assessed by JC-1 staining and mitochondrial gene transcription. Results: Sixteen hours of healthy human plasma exposure did not affect viability, however, hepatic gene-transcript levels decreased dramatically and dose-dependently within four hours of exposure. These changes were associated with early NF-kB signaling and a shift from mitochondrial energy metabolism towards glycolysis. Healthy human plasma-toxicity was associated with the dose-dependent presence of heat-resistant, albumin-bound and (partly) hydrophobic toxic compound(s). HepaRG cells cultured in BALs were partially protected from plasma-toxicity, which was mainly attributable to medium perfusion and/or 3D configuration applied during BAL culturing. The detrimental human plasma effects were reversible in BAL-cultured cells. Porcine ALF-plasma elicited mitotoxicity additional to the basal detrimental effect of porcine healthy plasma, which were only partially reversible. Conclusion: A specific fraction of human plasma reduces hepatic differentiation of HepaRG cultures, in association with early NF-κB activation. In addition, ALF-plasma elicits mitotoxic effects. These findings allow for a targeted approach in preventing plasma-induced cell damage.

show abstract

Section: Discussionsupporting

confidence: 91%

HepaRG-Progenitor Cell Derived Hepatocytes Cultured in Bioartificial Livers Are Protected from Healthy- and Acute Liver Failure-Plasma Induced Toxicity

Wenum¹,

Treskes²,

Adam³

et al. 2018

Cell Physiol Biochem

View full text Add to dashboard Cite

show abstract

“…Further evidence of a protective effect of three‐dimensional culture in a fulminant hepatic LF plasma environment can be seen in a study by Nagaki et al. which showed no toxic effects on primary hepatocytes (9). When primary hepatocytes are plated into monolayer culture, there are profound changes in gene expression relative to suspensions of human hepatocytes, including genes that are involved in cellular processes such as phase I/II metabolism, basolateral and canalicular transport systems, fatty acid and lipid metabolism, apoptosis, and proteasomal protein recycling (28).…”

Section: Discussionmentioning

confidence: 94%

“…Toxicity of plasma taken from patients with acute LF has been reported by a number of groups. However, only a few groups have attempted to assess its effects on cells grown in three‐dimensional cultures in the context of a bioartificial liver (9,10). Other studies that assess functionality of three‐dimensional hepatocyte cultures in putative bioartificial livers have perfused their systems with culture medium (11).…”

mentioning

confidence: 99%

Alginate‐encapsulated HepG2 Cells in a Fluidized Bed Bioreactor Maintain Function in Human Liver Failure Plasma

et al. 2009

View full text Add to dashboard Cite

Alginate-encapsulated HepG2 cells cultured in microgravity have the potential to serve as the cellular component of a bioartificial liver. This study investigates their performance in normal and liver failure (LF) human plasma over 6-8 h in a fluidized bed bioreactor. After 8 days of microgravity culture, beads containing 1.5 x 10(9) cells were perfused for up to 8 h at 48 mL/min with 300 mL of plasma. After exposure to 90% LF plasma, vital dye staining showed maintained cell viability, while a 7% increase in lactate dehydrogenase activity indicated minimal cell damage. Glucose consumption, lactate production, and a 4.3-fold linear increase in alpha-fetoprotein levels were observed. Detoxificatory function was demonstrated by quantification of bilirubin conjugation, urea synthesis, and Cyp450 1A activity. These data show that in LF plasma, alginate-encapsulated HepG2 cells can maintain viability, and metabolic, synthetic, and detoxificatory activities, indicating that the system can be scaled-up to form the biological component of a bioartificial liver.

show abstract

Proliferation of mouse liver stem/progenitor cells induced by plasma from patients with acute liver failure is modulated by P2Y2 receptor-mediated JNK activation

et al. 2013

View full text Add to dashboard Cite

show abstract

Effects of plasma from patients with fulminant hepatic failure on function of primary rat hepatocytes in three‐dimensional culture

Abstract: These findings demonstrated that FHF plasma is not toxic to rat hepatocyte spheroids and that hepatocyte spheroids have potential use in the development of a bioartificial liver.

Cited by 3 publications

References 30 publications

HepaRG-Progenitor Cell Derived Hepatocytes Cultured in Bioartificial Livers Are Protected from Healthy- and Acute Liver Failure-Plasma Induced Toxicity

HepaRG-Progenitor Cell Derived Hepatocytes Cultured in Bioartificial Livers Are Protected from Healthy- and Acute Liver Failure-Plasma Induced Toxicity

Alginate‐encapsulated HepG2 Cells in a Fluidized Bed Bioreactor Maintain Function in Human Liver Failure Plasma

Proliferation of mouse liver stem/progenitor cells induced by plasma from patients with acute liver failure is modulated by P2Y2 receptor-mediated JNK activation

Contact Info

Product

Resources

About