Urea Synthesis and Cyclosporin a Biotransformation in a Laboratory Scale Hepatocyte Bioreactor Model

Farghali, Hassan; D, Lincová; Gaier, Norbert; Kmoníèková, Eva; Kameníková, L; Canová, Nikolina Kutinová; Vı́tek, Libor

doi:10.1016/s1043661802002347

Cited by 3 publications

(3 citation statements)

References 13 publications

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“…Drug metabolism is preserved in rat primary hepatocytes cultured in WE [Wu et al, ; Takeba et al, ]. Gluconeogenesis and urea cycle—hepatocyte‐specific functions—are also maintained in WE [Farghali et al, ; Dabos et al, ]. These findings clearly indicate that WE is suitable for hepatocyte culture without loss of hepatocyte‐specific functions.…”

Section: Discussionmentioning

confidence: 99%

Transcription Factors and Medium Suitable for Initiating the Differentiation of Human‐Induced Pluripotent Stem Cells to the Hepatocyte Lineage

Tomizawa

Shinozaki

Motoyoshi

et al. 2016

J of Cellular Biochemistry

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Transcription factors and culture media were investigated to determine the condition to initiate the differentiation of human-induced pluripotent stem (iPS) cells most efficiently. The expression of genes in human adult liver was compared with that in 201B7 cells (iPS cells) using cDNA microarray analysis. Episomal plasmids expressing transcription factors were constructed. 201B7 cells were transfected with the episomal plasmids and cultured in ReproFF (feeder-free media maintaining pluripotency), Leibovitz-15 (L15), William's E (WE), or Dulbecco's modified Eagle medium/Nutrient F-12 Ham (DF12) for 7 days. RNA was isolated and subjected to real-time quantitative PCR to analyze the expression of alpha-feto protein (AFP) and albumin. cDNA microarray analysis revealed 16 transcription factors that were upregulated in human adult liver relative to that in 201B7 cells. Episomal plasmids expressing these 16 genes were transfected into 201B7 cells. CCAAT/enhancer-binding protein alpha (CEBPA), CCAAT/enhancer-binding protein beta (CEBPB), forkhead box A1 (FOXA1), and forkhead box A3 (FOXA3) up-regulated AFP and down-regulated Nanog. These four genes were further analyzed. The expression of AFP and albumin was the highest in 201B7 cells transfected with the combination of CEBPA, CEBPB, FOXA1, and FOXA3 and cultured in WE. The combination of CEBPA, CEBPB, FOXA1, and FOXA3 was suitable for 201B7 cells to initiate differentiation to the hepatocyte lineage and WE was the most suitable medium for culture after transfection. J. Cell. Biochem. 117: 2001-2009, 2016. © 2016 Wiley Periodicals, Inc.

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Section: Discussionmentioning

confidence: 99%

Transcription Factors and Medium Suitable for Initiating the Differentiation of Human‐Induced Pluripotent Stem Cells to the Hepatocyte Lineage

Tomizawa

Shinozaki

Motoyoshi

et al. 2016

J of Cellular Biochemistry

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“…Rat primary hepatocytes are reportedly capable of maintaining drug metabolism in WE [ 31 , 32 ]. Gluconeogenesis and urea synthesis are also preserved in hepatocytes cultured in WE [ 33 , 34 ]. These studies demonstrate that WE is the most suitable medium for the functional maintenance of cultured hepatocytes, and their results are consistent with those of the present study.…”

Section: Discussionmentioning

confidence: 99%

Improved Survival and Initiation of Differentiation of Human Induced Pluripotent Stem Cells to Hepatocyte-Like Cells upon Culture in William’s E Medium followed by Hepatocyte Differentiation Inducer Treatment

et al. 2016

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BackgroundHepatocyte differentiation inducer (HDI) lacks both glucose and arginine, but is supplemented with galactose and ornithine, and is added together with other reagents such as apoptosis inhibitor and oncostatin M. Although human induced pluripotent stem (iPS) cells initiate hepatocyte differentiation, most die within 7 days. In this study, we investigated both HDI and conventional media for their potential to improve cell survival.Materials and Methods201B7 iPS cells were cultured in conventional media. This consisted of three cycles of 5-day culture in William’s E (WE) medium, followed by a 2-day culture in HDI.ResultsExpression levels of α-feto protein (AFP) were higher in cells cultured in WE and in Dulbecco’s Modified Eagle’s Medium/Nutrient F-12 Ham (DF12). 201B7 cells expressed the highest AFP and albumin (ALB) when cultured in HDI for 2 days following 7-day culture in WE. After three cycles of 5-day culture in WE followed by 2 days in HDI, 201B7 cells expressed AFP and ALB 54 ± 2.3 (average ± standard deviation) and 73 ± 15.1 times higher, respectively, than those cultured in ReproFF (feeder-free condition).Conclusion201B7 cells survived culture in WE for 7 days followed HDI for 2 days. After three cycles of culture under these conditions, hepatocyte differentiation was enhanced, as evidenced by increased AFP and ALB expression.

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“…We have routinely used immobilised hepatocytes in agarose gel threads as a prototype of a laboratory-scale hepatocyte bioreactor model (28,29). Such a model is suitable for studying many general aspects of hepatocyte biology and pathobiology.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a Flat Membrane Hepatocyte Bioreactor for Pharmacotoxicological Applications: Evidence that Inhibition of Spontaneously Produced Nitric Oxide Improves Cell Functionality

Canová

Kmoníčková

et al. 2004

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A laboratory-scale bioreactor was re-evaluated, with the aim of improving its use for the perfused culture of rat hepatocytes. In contrast to conventional culture systems, the flat membrane bioreactor (FMB) showed good functionality and biochemical competence during 2–3 days. Hepatocytes cultured in the FMB, specifically in a “sandwich” configuration, were functionally stable, as shown by a high rate of urea biosynthesis after challenge with NH4Cl, a low alanine-aminotransferase leakage and suppressed spontaneous nitric oxide (NO) production. Moreover, the time-course of the disappearance of cyclosporin A (CsA) from the perfusate demonstrated the high biotransformation capacity of cells in the FMB. The effect of CsA on the modulation of urea and spontaneous NO production demonstrated flexibility, in that minor changes could be observed at diverse time intervals and in a non-destructive way. The monitoring of nitrite levels during various steps of isolation and culture suggested that spontaneously produced NO has a negative impact on hepatocyte metabolic and functional integrity. In spite of the sophisticated techniques that are being used for the preparation of bioreactors, with hepatocytes surviving for longer periods, our data have shed light on some factors that could be important for the successful use of similar models for pharmacotoxicological and other biomedical applications.

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Urea Synthesis and Cyclosporin a Biotransformation in a Laboratory Scale Hepatocyte Bioreactor Model

Cited by 3 publications

References 13 publications

Transcription Factors and Medium Suitable for Initiating the Differentiation of Human‐Induced Pluripotent Stem Cells to the Hepatocyte Lineage

Transcription Factors and Medium Suitable for Initiating the Differentiation of Human‐Induced Pluripotent Stem Cells to the Hepatocyte Lineage

Improved Survival and Initiation of Differentiation of Human Induced Pluripotent Stem Cells to Hepatocyte-Like Cells upon Culture in William’s E Medium followed by Hepatocyte Differentiation Inducer Treatment

Evaluation of a Flat Membrane Hepatocyte Bioreactor for Pharmacotoxicological Applications: Evidence that Inhibition of Spontaneously Produced Nitric Oxide Improves Cell Functionality

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