Dynamic Seeding and<i>in Vitro</i>Culture of Hepatocytes in a Flow Perfusion System

Kim, Stephen S.; Sundback, Cathryn A.; Kaihara, Satoshi; Benvenuto, Mark; Kim, Byung Soo; Mooney, David J.; Vacanti, Joseph P.

doi:10.1089/107632700320874

Cited by 129 publications

(73 citation statements)

References 24 publications

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“…4 By using perfusion systems, cells can adequately be supplied with nutrients and oxygen, while wastes are removed permanently. 5,6 The improvement of hepatocytes' proliferation and metabolism by a constant perfusion flow in a microfluidic reactor was shown previously. 5 Nevertheless, a strong flow does not reflect the in vivo situation as a hepatocyte in the liver is not in direct contact to blood circulation for endothelial cells and the space of Disse acting as filters to the hepatocytes.…”

Section: Introductionmentioning

confidence: 77%

On-chip three-dimensional cell culture in phaseguides improves hepatocyte functions in vitro

et al. 2015

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The in vitro study of liver functions and liver cell specific responses to external stimuli deals with the problem to preserve the in vivo functions of primary hepatocytes. In this study, we used the biochip OrganoPlate TM (MIMETAS) that combines different advantages for the cultivation of hepatocytes in vitro: (1) the perfusion flow is achieved without a pump allowing easy handling and placement in the incubator; (2) the phaseguides allow plating of matrix-embedded cells in lanes adjacent to the perfusion flow without physical barrier; and (3) the matrix-embedding ensures indirect contact of the cells to the flow. In order to evaluate the applicability of this biochip for the study of hepatocyte's functions, Matrigel TM -embedded HepG2 cells were cultured over three weeks in this biochip and compared to a static Matrigel culture (3D) and a monolayer culture (2D). Chip-cultured cells grew in spheroid-like structures and were characterized by the formation of bile canaliculi and a high viability over 14 days. Hepatocyte-specific physiology was achieved as determined by an increase in albumin production. Improved detoxification metabolism was demonstrated by strongly increased cytochrome P450 activity and urea production. Additionally, chip-cultured cells displayed increased sensitivity to acetaminophen. Altogether, the OrganoPlate seems to be a very useful alternative for the cultivation of hepatocytes, as their behavior was strongly improved over 2D and static 3D cultures and the results were largely comparable and partly superior to the previous reports on biochip-cultured hepatocytes. As for the low technical needs, this platform has the appearance of being highly applicable for further studies of hepatocytes' responses to external stimuli. V C 2015 AIP Publishing LLC. [http://dx

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Section: Introductionmentioning

confidence: 77%

On-chip three-dimensional cell culture in phaseguides improves hepatocyte functions in vitro

et al. 2015

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“…Other dynamic scaffold seeding techniques are described in the tissue engineering literature. [38][39][40][41][42][43][44][45] In one such approach, the carrier scaffold is suspended in a spinner flask containing culture medium. The medium is then inoculated with cells that attach to the carrier over time.…”

Section: Discussionmentioning

confidence: 99%

A Rapid Seeding Technique for the Assembly of Large Cell/Scaffold Composite Constructs

Solchaga¹,

Tognana²,

Penick³

et al. 2006

Tissue Engineering

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These studies address critical technical issues involved in creating human mesenchymal stem cell (hMSC)/scaffold implants for cartilage repair. These issues include obtaining a high cell density and uniform spatial cell distribution within the scaffold, factors that are critical in the initiation and homogeneity of chondrogenic differentiation. For any given scaffold, the initial seeding influences cell density, retention, and spatial distribution within the scaffold, which eventually will affect the function of the construct. Here, we discuss the development of a vacuum-aided seeding technique for HYAFF ® -11 sponges which we compared to passive infiltration. Our results show that, under the conditions tested, hMSCs were quantitatively and homogeneously loaded into the scaffolds with 90+% retention rates after 24 h in perfusion culture with no negative effect on cell viability or chondrogenic potential. The retention rates of the vacuum-seeded constructs were at least 2 times greater than those of passively seeded constructs at 72 h. Histomorphometric analysis revealed that the core of the vacuum-seeded constructs contained 240% more cells than the core of passively infiltrated scaffolds. The vacuum seeding technique is safe, rapid, reproducible, and results in controlled quantitative cell loading, high retention, and uniform distribution.

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“…In the last two decades innumerous strategies for the culture of adult hepatocytes in combination with several types of 3D, highly porous polymeric matrices have been attempted (45)(46)(47)(48)(49). However, in the absence of vasculature, restriction in cell growth and function is common due to the limitations in nutrient and oxygen diffusion.…”

Section: Liver Bioengineeringmentioning

confidence: 99%