A Device to Measure the Oxygen Uptake Rate of Attached Cells: Importance in Bioartificial Organ Design

Foy, Brent D.; Rotem, Avi; Toner, Mehmet; Tompkins, Ronald G.; Yarmush, Martin L.

doi:10.1177/096368979400300609

Cited by 91 publications

(86 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We assumed the D values for oxygen and glucose in the medium and cell layer are the same values as those of water at 310 K and estimated them to be 2.10ϫ 10 −9 and 9.91ϫ 10 −10 m 2 / s for oxygen and glucose, respectively. The values of V max and K h for oxygen consumption by rat primary hepatocytes were found to be 2.0ϫ 10 −16 mol/ s cell and 4.05ϫ 10 −3 mol/ m 3 , respectively, based on the data obtained by Foy et al 18 Not finding any values in literature, we estimated the V max and K h values for glucose in the microwells to be one-sixth of the corresponding values for oxygen because aerobic respiration consumes 6 moles of oxygen per mole of glucose. …”

Section: -3mentioning

confidence: 64%

Superior oxygen and glucose supply in perfusion cell cultures compared to static cell cultures demonstrated by simulations using the finite element method

et al. 2011

View full text Add to dashboard Cite

Oxygen and glucose supply is one of the important factors for the growth and viability of the cells in cultivation of tissues, e.g., spheroid, multilayered cells, and three-dimensional tissue construct. In this study, we used finite element methods to simulate the flow profile as well as oxygen and glucose supply to the multilayered cells in a microwell array chip for static and perfusion cultures. The simulation results indicated that oxygen supply is more crucial than glucose supply in both static and perfusion cultures, and that the oxygen supply through the wall of the perfusion culture chip is important in perfusion cultures. Glucose concentrations decline with time in static cultures, whereas they can be maintained at a constant level over time in perfusion cultures. The simulation of perfusion cultures indicated that the important parameters for glucose supply are the flow rate of the perfusion medium and the length of the cell culture chamber. In a perfusion culture chip made of oxygen-permeable materials, e.g., polydimethylsiloxane, oxygen is hardly supplied via the perfusion medium, but mainly supplied through the walls of the perfusion culture chip. The simulation of perfusion cultures indicated that the important parameters for oxygen supply are the thickness of the flow channel and the oxygen permeability of the walls of the channel, i.e., the type of material and the thickness of the wall.

show abstract

Section: -3mentioning

confidence: 64%

Superior oxygen and glucose supply in perfusion cell cultures compared to static cell cultures demonstrated by simulations using the finite element method

et al. 2011

View full text Add to dashboard Cite

show abstract

“…Unfortunately, the Hypoxyprobe assay is not as effective in helping to discern if this intracellular pressure translates to hypoxic conditions for these cells since the literature indicates that pO 2 less than 10 mmHg are not necessarily hypoxic for hepatocytes maintained in vitro. More specifically, the O 2 tension reported for regular in vitro tissue cultures ranges between 2.3 and 5.3 mmHg, when the surrounding environment is maintained at 20% O 2 , [42][43][44] and has been shown to reach an intracellular pO 2 of only 11 mmHg after equilibrating the cell culture medium to 159 mmHg pO 2 . Even, the physiological range of O 2 tension in the natural liver is known to span from 5 to 90 mmHg.…”

Section: Discussionmentioning

confidence: 99%

The Effectiveness of a Novel Cartridge-Based Bioreactor Design in Supporting Liver Cells

Niu

Hammond

Coger

2009

Tissue Engineering Part A

View full text Add to dashboard Cite

There are a number of applications-ranging from temporary strategies for organ failure to pharmaceutical testing-that rely on effective bioreactor designs. The significance of these devices is that they provide an environment for maintaining cells in a way that allows them to perform key cellular and tissue functions. In the current study, a novel cartridge-based bioreactor was developed and evaluated. Its unique features include its capacity for cell support and the adaptable design of its cellular space. Specifically, it is able to accommodate functional and reasonably sized tissue (>2.0Â10 8 cells), and can be easily modified to support a range of anchorage-dependent cells. To evaluate its efficacy, it was applied to liver support in the current study. This involved evaluating the performance of rat primary hepatocytes within the unique cartridges in culture-sans bioreactor-and after being loaded within the novel bioreactor. Compared to collagen sandwich culture functional controls, hepatocytes within the unique cartridge design demonstrated significantly higher albumin production and urea secretion rates when cultured under dynamic flow conditions-reaching peak values of 170 AE 22 mg=10 6 cells=day and 195 AE 18 mg=10 6 cells=day, respectively. The bioreactor's effectiveness in supporting live and functioning primary hepatocytes is also presented. Cell viability at the end of 15 days of culture in the new bioreactor was 84 AE 18%, suggesting that the new design is effective in maintaining primary hepatocytes for at least 2 weeks in culture. Liver-specific functions of urea secretion, albumin synthesis, and cytochrome P450 activity were also assessed. The results indicate that hepatocytes are able to achieve good functional performance when cultured within the novel bioreactor. This is especially true in the case of cytochrome P450 activity, where by day 15 of culture, hepatocytes within the bioreactor reached values that were 56.6% higher than achieved by the collagen sandwich functional control cultures. The success of the novel cartridge-based bioreactor in supporting hepatocytes with good viability and functional performance suggests that it is an effective design for supporting anchorage-dependent cells.

show abstract

“…10 6 cells/cm 2 ) attachment rates to the culture carrier were drastically reduced and the LDH release of cells increased indicating cell damage. This could be due to a shortage of available oxygen in particular in the early phase of culture, since hepatocytes that have undergone an enzymatic cell isolation protocol do have an elevated need for oxygen [Foy et al, 1994]. Therefore, as suggested by other authors, we used a 40% oxygen atmosphere [Yanagi et al, 1998].…”

Section: Discussionmentioning

confidence: 99%

High Density Culturing of Porcine Hepatocytes Immobilized on Nonwoven Polyurethane-Based Biomatrices

Pahernik¹,

Thasler²,

Doser

et al. 2001

Cells Tissues Organs

View full text Add to dashboard Cite

Objective: Hepatocytes are increasingly used as functional units in bioartificial liver devices. The objective of the present study was to investigate the feasibility of cul-turing porcine hepatocytes in high density on a novel polyurethane-based nonwoven three-dimensional matrix. We investigated (1) the optimal cell density within this culture configuration, (2) the maintenance of liver-specific morphology and cell functions over long-term periods and (3) the necessity to apply an additional extra-cellular matrix component (collagen gel). Methods: Non-woven polyurethane matrices were manufactured by a specially developed fiber extrusion technology. Pig he-patocytes were cultured at various cell densities of 0.1, 0.25, 0.5, 0.75, 1 and 2 ! 10 6 cells/cm 2 on three-dimensional networks of nonwoven polyurethane matrices and cell adhesion as well as functional parameters (DNA of nonattached/attached cells, lactate dehydrogenase release and cytochrome P450 activity) were determined. To assess the performance of cells within this configuration albumin and urea excretion was measured over 8 days. The potentially beneficial effect of an additional extracellular matrix configuration was evaluated by comparing the average albumin synthesis in groups of identical cell numbers. Results: The optimal cell density in this three-dimensional culture configuration was 1 ! 10 6 cells/cm 2. The functional capacity of hepatocytes was stable for 8 days at an average level of 53.7 B 5.6 ng/h/Ìg DNA and of 1.8 B 0.14 Ìg/h/Ìg DNA for albumin and urea excretion, respectively. The supplementation of an extracellular matrix configuration did not improve functional activity of cells. Average albumin synthesis was 35.6 ng/h/Ìg DNA (28.7, 42.8) and 32.7 ng/h/Ìg DNA (23.4, 49.2) for collagen-immobilized and control cul-High-Density Culturing of Hepatocytes Cells Tissues Organs 2001;168:170-177 171 tures, respectively. Conclusion: The results of the study indicate that nonwoven polyurethane sheets supply a biocompatible support structure for functionally active high density cultures. Thus, nonwoven polyurethane matrices should be further investigated on with respect to their role in the development, optimization and design of bioartificial liver systems.

show abstract

A Device to Measure the Oxygen Uptake Rate of Attached Cells: Importance in Bioartificial Organ Design

Cited by 91 publications

References 28 publications

Superior oxygen and glucose supply in perfusion cell cultures compared to static cell cultures demonstrated by simulations using the finite element method

Superior oxygen and glucose supply in perfusion cell cultures compared to static cell cultures demonstrated by simulations using the finite element method

The Effectiveness of a Novel Cartridge-Based Bioreactor Design in Supporting Liver Cells

High Density Culturing of Porcine Hepatocytes Immobilized on Nonwoven Polyurethane-Based Biomatrices

Contact Info

Product

Resources

About