The Effect of Oxygen Transport Resistances on the Viability and Functions of Isolated Rat Hepatocytes

Abstract

The treatment of fulminant hepatic failure with a bioartificial liver support device relies on the possibility of replacing the detoxification and synthetic functions of the injured liver for as long as needed for patient recovery. In spite of progress in cell culture techniques, the effective use of isolated hepatocytes in liver support devices is currently hampered by a lack of information on the metabolic factors limiting long term hepatocyte culture. In this paper, we report our investigation on the effects of oxygen transport resistances on the viability and functions of isolated rat hepatocytes cultured on collagen coated Petri dishes. Detoxification and synthetic functions of the hepatocytes were studied with respect to ammonia and phenolsulphonphthalein elimination and urea synthesis. Lower resistances to oxygen transport favored hepatocyte survival. The isolated hepatocytes synthesized urea at rates that decreased as the resistance to oxygen transport increased. The rate at which urea was synthesized also decreased during the culture. Neither PSP, nor ammonia elimination rate was greatly affected by increasing oxygen transport resistances and remained rather constant up to a week of culture.

In vitro evaluation of a novel bioreactor based on an integral oxygenator and a spirally wound nonwoven polyester matrix for hepatocyte culture as small aggregates

“…In addition, we wanted to culture hepatocytes as small aggregates, known to maintain many of the cytoarchitectural characteristics found in vivo and to exhibit higher and more prolonged functional activity than hepatocytes cultured in monolayers (25)(26)(27). Special attention was paid to optimal oxygenation of the cells, since oxygen plays an important role in hepatocyte attachment (28) and function (29,30). Therefore, the bioreactor was equipped with an integrated oxygenator.…”

Model-Based Optimization of Scaffold Geometry and Operating Conditions of Radial Flow Packed-Bed Bioreactors for Therapeutic Applications

“…The attention was focused on oxygen for its importance to cell metabolism [4][5][6][7][8]. Schemes of an rPBB with opposite perfusion flows are reported in Figure 1.…”

“…Cell culture in the presence of controlled physiological concentrations and supply of nutrients and dissolved oxygen is a basic bioreactor requirement to ensure cell survival and to make cells differentiate to a given phenotype. In particular, oxygen is acknowledged a major limiting role to the development of engineered substitutes mimicking natural tissues [4][5][6][7][8]. Metabolic soluble species are generally supplied in the culture medium.…”

Nanometric self-assembling peptide layers maintain adult hepatocyte phenotype in sandwich cultures

“…Furthermore, it was shown as early as in 1968 that commonly used medium depths of 2-5 mm in Petri dishes rapidly produced hypoxic conditions when hepatocytes respired at their physiological rate [22]. Therefore, because plastic walls and culture medium are efficient barriers of oxygen diffusion, it is important to create a system in which a physiological oxygen supply is maintained [23,24]. …”