Metabolic Patterning on a Chip: Towards in vitro Liver Zonation of Primary Rat and Human Hepatocytes

Kang, Young Bok Abraham; Eo, Jinsu; Mert, Safak; Yarmush, Martin L.; Usta, O. Berk

doi:10.1038/s41598-018-27179-6

Cited by 99 publications

(112 citation statements)

References 54 publications

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“…Further, the specific design of our biochip enables to selectively close the small pillars allowing the development of a metabolic zonation, which is a critical issue in liver‐like models (Y. B. Kang, Eo, Mert, Yarmush, & Usta, ). The CFD simulations were carried out to correlate the effects of the culture microenvironment to tissue modification over time.…”

Section: Resultsmentioning

confidence: 99%

A three‐dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity

Corrado

Gregorio

Imparato

et al. 2019

Biotech & Bioengineering

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In this study, we propose the design and fabrication of a liver system on a chip. We first chose the most suitable three‐dimensional liver‐like model between cell spheroids and microtissue precursors, both based on the use of hepatocellular carcinoma cells (HepG2) to provide proof‐of‐concept data. Spheroids displayed high cell density but low expression of the typical hepatic biomarkers, whereas microtissue precursors showed stable viability and function over the entire culture time. The two liver‐like models were compared in terms of cell viability, function, metabolism, and the P‐glycoprotein 1 (P‐gp) transport‐protein expression with the microtissue precursors showing the best performance. Thus, we cultured them into a microfluidic biochip featured with three parallel channels shaped to mimic the hepatic sinusoids. To assess the detoxification potential of the microtissue‐loaded biochip we challenged it with a model molecule (ethanol) at different concentrations and time points. Ethanol cytotoxicity was detected by a noninvasive measurement of cell viability based on cell autofluorescence. As expected, a dose‐dependent decrease of albumin and urea secretion was observed in the ethanol‐treated samples. We believe that the described totally human‐derived platform, suitable for integration into a multiorgan microfluidic system, can provide a consistent innovative platform for drug development and toxicity studies.

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Section: Resultsmentioning

confidence: 99%

A three‐dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity

Corrado

Gregorio

Imparato

et al. 2019

Biotech & Bioengineering

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“…The design and fabrication of the gradient‐generating MPOC device were explained in detail in our previous study (Kang et al, ). Briefly, it is composed of a gradient generator (width [w] × height [h]: 75 × 200 µm) and a cell culture chamber (wiggle shape, w × h × length [l]: 10,000 × 200 × 1,700 µm), in addition to inlets and outlets for media flow and cell seeding.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, it is composed of a gradient generator (width [w] × height [h]: 75 × 200 µm) and a cell culture chamber (wiggle shape, w × h × length [l]: 10,000 × 200 × 1,700 µm), in addition to inlets and outlets for media flow and cell seeding. All devices used in this study were fabricated by PDMS replica molding from a SU‐8 template made with the help of photolithography technology, as previously described (Kang et al, ; McCarty et al, ). The PDMS microfluidic device was bonded on a glass microscope slide (w × l: 25 × 75 mm; Thermo Fisher Scientific, Grand Island, NY) after oxygen plasma treatment.…”

Section: Methodsmentioning

confidence: 99%

“…Primary rat hepatocytes were freshly isolated from adult female Lewis rats (Charles River Laboratories, MA) according to the protocol #2011N000111 approved by the Institutional Animal Care and Use Committee (Dunn, Tompkins, & Yarmush, ; Seglen, ). Approximately 3.1–4.1 × 10 4 hepatocytes with 90–95% viability were cultured in the cell culture chamber of the device that was coated with 50 μg/ml fibronectin (Sigma‐Aldrich, St. Louis, MO) for 20–40 min at 37°C and 10% CO 2 as described previously (Kang et al, ; McCarty et al, ). The cells were seeded and cultured at 37°C and 10% CO 2 in Dulbecco's modified eagle's medium (DMEM; Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum (Sigma‐Aldrich), 0.5 U/ml insulin, 7 ng/ml glucagon, 20 ng/ml epidermal growth factor, 7.5 μg/ml hydrocortisone, 200 U/ml penicillin, 200 μg/ml streptomycin, and 50 μg/ml gentamycin.…”

Section: Methodsmentioning

confidence: 99%

“…In our previous studies, we developed a microfluidic platform, the “Metabolic Patterning on a Chip (MPOC) device”, and successfully demonstrated in vitro zonation of primary rat and human hepatocytes via this platform. The MPOC device was able to create actively‐controlled gradients of inducers or hormones, and thus enforce the creation of metabolic patterns across microscopic tissues (Kang, Eo, Mert, Yarmush, & Usta, ; McCarty, Usta, & Yarmush, ).…”

Section: Introductionmentioning

confidence: 99%

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Progressive hypoxia‐on‐a‐chip: An in vitro oxygen gradient model for capturing the effects of hypoxia on primary hepatocytes in health and disease

Kang

Bulutoglu

et al. 2019

Biotech & Bioengineering

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Oxygen is vital to the function of all tissues including the liver and lack of oxygen, that is, hypoxia can result in both acute and chronic injuries to the liver in vivo and ex vivo.Furthermore, a permanent oxygen gradient is naturally present along the liver sinusoid, which plays a role in the metabolic zonation and the pathophysiology of liver diseases. Accordingly, here, we introduce an in vitro microfluidic platform capable of actively creating a series of oxygen concentrations on a single continuous microtissue, ranging from normoxia to severe hypoxia. This range approximately captures both the physiologically relevant oxygen gradient generated from the portal vein to the central vein in the liver, and the severe hypoxia occurring in ischemia and liver diseases. Primary rat hepatocytes cultured in this microfluidic platform were exposed to an oxygen gradient of 0.3-6.9%. The establishment of an ascending hypoxia gradient in hepatocytes was confirmed in response to the decreasing oxygen supply.The hepatocyte viability in this platform decreased to approximately 80% along the hypoxia gradient. Simultaneously, a progressive increase in accumulation of reactive oxygen species and expression of hypoxia-inducible factor 1α was observed with increasing hypoxia. These results demonstrate the induction of distinct metabolic and genetic responses in hepatocytes upon exposure to an oxygen (/hypoxia) gradient.This progressive hypoxia-on-a-chip platform can be used to study the role of oxygen and hypoxia-associated molecules in modeling healthy and injured liver tissues. Its use can be further expanded to the study of other hypoxic tissues such as tumors as well as the investigation of drug toxicity and efficacy under oxygen-limited conditions. K E Y W O R D S hepatocytes, HIF, hypoxia, microfluidics, oxygen gradient, ROS

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Temporal Dynamics of Metabolic Acquisition in Grafted Engineered Human Liver Tissue

et al. 2022

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Liver disease affects millions globally, and end‐stage liver failure is only cured by organ transplant. Unfortunately, there is a growing shortage of donor organs as well as inequitable access to transplants across populations. Engineered liver tissue grafts that supplement or replace native organ function can address this challenge. While engineered liver tissues have been successfully engrafted previously, the extent to which these tissues express human liver metabolic genes and proteins remains unknown. Here, it is built engineered human liver tissues and characterized their engraftment, expansion, and metabolic phenotype at sequential stages post‐implantation by RNA sequencing, histology, and host serology. Expression of metabolic genes is observed at weeks 1–2, followed by the cellular organization into hepatic cords by weeks 4–9.5. Furthermore, grafted engineered tissues exhibited progressive spatially restricted expression of critical functional proteins known to be zonated in the native human liver. This is the first report of engineered human liver tissue zonation after implantation in vivo, which can have important translational implications for this field.

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Metabolic Patterning on a Chip: Towards in vitro Liver Zonation of Primary Rat and Human Hepatocytes

Cited by 99 publications

References 54 publications

A three‐dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity

A three‐dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity

Progressive hypoxia‐on‐a‐chip: An in vitro oxygen gradient model for capturing the effects of hypoxia on primary hepatocytes in health and disease

Temporal Dynamics of Metabolic Acquisition in Grafted Engineered Human Liver Tissue

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