Techniques for measurement of oxygen consumption rates of hepatocytes during attachment and post-attachment.
Three techniques for measuring oxygen consumption rate (OCR) of cultured cells relevant to the development of bioartificial liver devices are reported. In an oxystat apparatus, HepG2 cells immobilised on Cytodex 3 microcarriers at a concentration of 10(6) cells ml-1 had a mean OCR of 0.7 nmol s-1/10(6) cells. The OCR decreased with increasing cell density, a characteristic previously reported for other cell lines. Rat hepatocytes immobilised on single collagen layers in a flow cell and challenged with ammonia had a mean OCR of 0.59 nmol s-1/10(6) cells. A novel two-compartment oxystat system was used to determine the OCR of rat hepatocytes during the attachment phase. OCR declined from 1.0 nmol s-1/10(6) immediately after seeding to 0.7 nmol s-1/10(6) cells at nine hours. The low OCR for HepG2 reflects loss of certain oxygen dependent metabolic pathways. The OCR measured for rat hepatocytes during and post-attachment are significantly higher than those reported elsewhere and have major implications for the development of bioartificial liver devices.
Smith
M D
MD
Bioengineering Unit, University of Strathclyde, Glasgow, Scotland, UK.
Smirthwaite
A D
AD
Cairns
D E
DE
Cousins
R B
RB
Gaylor
J D
JD
D000818:Animals; D001187:Artificial Organs / Q000592:standards; D001709:Biotechnology; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry / Q000378:metabolism; D003911:Dextrans / Q000737:chemistry / Q000378:metabolism; D005782:Gels; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008113:Liver Neoplasms / Q000473:pathology; D008297:Male; D008863:Microspheres; D010101:Oxygen Consumption / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014407:Tumor Cells, Cultured
D003911:Dextrans; D005782:Gels; C052839:Cytodex; D003094:Collagen