Porcine hepatocyte–Kupffer cell co‐culture as an <i>in vitro</i> model for testing the efficacy of anti‐inflammatory substances

Mátis, Gábor; Kulcsár, Anna; Petrilla, Janka; Talapka, P; Neogrády, Zs.

doi:10.1111/jpn.12547

Cited by 9 publications

(14 citation statements)

References 16 publications

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“…Aside from its primary function as an energy source for colonocytes, it is a strong mitosis promoter and a differentiation agent for intestinal epithelial cells [8], as it acts as a histone deacetylase (HDAC) inhibitor [9]. It showed also in vitro positive effects on hepatocytes [10]. Moreover, it has a strong antibacterial activity against both Gram-negative and Gram-positive pathogens [11] and therefore proves to be a valid aid for gut health maintenance.…”

Section: Introductionmentioning

confidence: 99%

Effects of Tributyrin Supplementation on Growth Performance, Insulin, Blood Metabolites and Gut Microbiota in Weaned Piglets

Sotira

Dell’Anno

Caprarulo

et al. 2020

Animals

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The aim of this study was to investigate the effects of tributyrin supplementation on the production traits, the main metabolic parameters and gut microbiota in weaned piglets. One hundred and twenty crossbred piglets (Large White × Landrace) were randomly divided into two experimental groups (six pens each; 10 piglets per pen): the control group (CTRL), that received a basal diet, and the tributyrin group (TRIB) that received the basal diet supplemented with 0.2% tributyrin. The experimental period lasted 40 days. Production traits were measured at days 14, 28 and 40. A subset composed of 48 animals (n = 4 for each pen; n = 24 per group) was considered for the evaluation of serum metabolic parameters and hair cortisol by enzyme-linked immunosorbent assay (ELISA), and faecal microbiota by real-time polymerase chain reaction (PCR). Our results showed that the treatment significantly increased body weight (BW) at day 28 and day 40 (p = 0.0279 and p = 0.0006, respectively) and average daily gain (ADG) from day 28 to day 40 (p = 0.046). Gain to feed ratio (G:F) was significantly higher throughout the experimental period (p = 0.049). Even if the serum parameters were in the physiological range, albumin, albumin/globulin (A/G) ratio, glucose and high-density lipoproteins (HDL) fraction were significantly higher in the TRIB group. On the contrary, tributyrin significantly decreased the urea blood concentration (p = 0.0026), which was correlated with lean gain and feed efficiency. Moreover, serum insulin concentration, which has a regulatory effect on protein and lipid metabolism, was significantly higher in the TRIB group (p = 0.0187). In conclusion, this study demonstrated that tributyrin can be considered as a valid feed additive for weaned piglets.

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Section: Introductionmentioning

confidence: 99%

Effects of Tributyrin Supplementation on Growth Performance, Insulin, Blood Metabolites and Gut Microbiota in Weaned Piglets

Sotira

Dell’Anno

Caprarulo

et al. 2020

Animals

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“…A monolayer hepatocyte-Kupffer cell and a double-layered enterohepatic co-culture have been also established recently by our research group from swine [13,19], but to the best of our knowledge, no similar avian cell cultures were available until now. From chickens, mainly tumorigenic cell lines, such as the Chicken Hepatocellular Carcinoma Cell Line (indicated as LMH cells) [20,21] or embryonic liver cell cultures [22] were used.…”

Section: Discussionmentioning

confidence: 99%

“…The newly prepared chicken cell cultures enable studies concerning the specific role of parenchymal and NP cells as the main liver cell fraction, and the hepatocyte-NP cell co-culture model can mimic various inflammatory states by setting different cell type ratios. The applied ratio of 6:1 (hepatocytes to NP cells) refers to a milder hepatic inflammation with moderate intrahepatic macrophage migration [13]. On this co-culture, the interaction of the inflammatory and stress response can be studied, including molecular alterations of cell function, such as the pro-and anti-inflammatory cytokine production and the redox homeostasis of the cultured liver cells.…”

Section: Discussionmentioning

confidence: 99%

Effects of Acute Heat Stress on a Newly Established Chicken Hepatocyte—Nonparenchymal Cell Co-Culture Model

Mackei

Molnár

Nagy

et al. 2020

Animals

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Heat stress is one of the most important issues in broiler flocks impairing animal health and productivity. On a cellular level, excess heat exposure can trigger heat shock response acting for the restoration of cell homeostasis by several mechanisms, such as affecting heat shock protein synthesis, redox homeostasis and pro-inflammatory cytokine production. The major aim of this study was to establish a novel avian hepatocyte—nonparenchymal cell co-culture as a model for investigating the cellular effects of heat stress and its interaction with inflammation in chicken liver. Cell fractions were isolated by differential centrifugation from a freshly perfused chicken liver, and hepatocyte mono-cultures as well as hepatocyte–nonparenchymal cell co-cultures (with cell ratio 6:1, hepatocytes to nonparenchymal cells, mimicking a milder hepatic inflammation) were prepared. Isolated and cultured cells were characterized by flow cytometry and immunocytochemistry applying hepatocyte- and macrophage-specific antibodies. Confluent cell cultures were exposed to 43 °C temperature for 1 or 2 h, while controls were cultured at 38.5 °C. The metabolic activity, LDH enzyme activity, reactive oxygen species (H2O2) production, extracellular concentration of heat shock protein 70 (HSP70), and that of the pro-inflammatory cytokines interleukin (IL-)6 and IL-8 were assessed. Shorter heat stress applied for 1 h could strongly influence liver cell function by significantly increasing catabolic metabolism and extracellular H2O2 release, and by significantly decreasing HSP70, IL-6, and IL-8 production on both cell culture models. However, all these alterations were restored after 2 h heat exposure, indicating a fast recovery of liver cells. Hepatocyte mono-cultures and hepatocyte—nonparenchymal cell co-cultures responded to heat stress in a similar manner, but the higher metabolic rate of co-cultured cells may have contributed to a better capability of inflamed liver cells for accommodation to stress conditions. In conclusion, the established new primary cell culture models provide suitable tools for studying the hepatic inflammatory and stress response. The results of this study highlight the impact of short-term heat stress on the liver in chickens, underline the mediatory role of oxidative stress in acute stress response, and suggest a fast cellular adaptation potential in liver cells.

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“…Generally, conventional in vitro culture systems are known to lack hepatocyte functions; therefore, it is necessary to establish and make available a functional culture system that maintains their specific function. Many studies have demonstrated the maintenance and differentiation of hepatocytes cultured with a variety of non-hepatic cells [21][22][23]. The cell-associated and soluble factors were shown to keep the hepatocytes morphologically and functionally differentiated [24][25][26].…”

Section: Discussionmentioning

confidence: 99%

Flow-Based Three-Dimensional Co-Culture Model for Long-Term Hepatotoxicity Prediction

2019

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We developed concave microwell arrays to establish a size-controllable 3-D co-culture liver model for in vitro drug toxicity testing, to predict hepatotoxicity. The interaction of hepatocytes with hepatic stellate cells (HSCs) was investigated by co-culturing primary 3-D hepatocyte spheroids and HSCs (heterosphere), using 3-D liver-on-a-chip. The effect of HSCs was investigated during spheroid formation; they were involved in controlling the organization of spheroidal aggregates and the formation of tight cell–cell contacts. Scanning electron microscopy (SEM) images showed that co-cultured spheroids with smoother surfaces in the flow chip aggregated more tightly and rapidly, compared to mono-cultured spheroids, until 13 days. Metabolic function analysis revealed that heterospheres secreted 40% more albumin and urea than hepatospheres on day 13. Additionally, an acetaminophen (AAP) and isoniazid (INH) concentration-dependent increase in CYP3A4 expression was detected in the 3-D cultures, and an increase in Lactate dehydrogenase (LDH) release after AAP and INH treatment was observed. CYP1A2, Mrp1 and UGT1A5 mRNA expression levels in the heterospheres and hepatospheres were evaluated from days 3 to 13. To examine the potential for toxicity testing in the flow-conditioned culture of the heterospheres, we evaluated cytotoxicity using the endpoint LDH release in the heterospheres and hepatospheres. IC50 values for AAP and INH after 24 h of exposure were calculated from the dose–response curves of the compounds. Flow-conditioned heterosphere culture results suggest that it may be suitable for long-term culture and cytotoxicity testing. Thus, our co-culture system closely resembles the in vivo environment and allows long-term in vitro hepatotoxicity prediction.

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Porcine hepatocyte–Kupffer cell co‐culture as an in vitro model for testing the efficacy of anti‐inflammatory substances

Cited by 9 publications

References 16 publications

Effects of Tributyrin Supplementation on Growth Performance, Insulin, Blood Metabolites and Gut Microbiota in Weaned Piglets

Effects of Tributyrin Supplementation on Growth Performance, Insulin, Blood Metabolites and Gut Microbiota in Weaned Piglets

Effects of Acute Heat Stress on a Newly Established Chicken Hepatocyte—Nonparenchymal Cell Co-Culture Model

Flow-Based Three-Dimensional Co-Culture Model for Long-Term Hepatotoxicity Prediction

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