Real-time monitoring of lactate extrusion and glucose consumption of cultured cells using a lab-on-valve system.
Microsequential injection (microST) provides microfluidic operations that are ideally suited for cellular function studies and as a means of validating targets for drug discovery. MicroSI carried out within the lab-on-valve (LOV) manifold, is an ideal platform for spectroscopic studies on living cells that are grown on microcarrier beads and kept thermostated while their metabolism is probed in real-time. In this paper a microbioreactor is integrated into the LOV manifold allowing measurement of cellular lactate extrusion and glucose consumption rates of a cell culture that is automatically renewed prior to each measurement. Glucose consumption and lactate extrusion are monitored using NAD-linked enzymatic assays. The microSI-LOV setup has demonstrated a linear analysis range of 0.05-1.00 mM for lactate and 0.1-5.6 mM for glucose. These assays were conducted in a serial fashion requiring 3 microL of cellular perfusate and 10 s for glucose determination and 30 s for the lactate assay. Overall waste generated per lactate/glucose assay is < 200 microL. This work was performed using two different transfected hepatocyte cell lines, which adhere to Cytopore microcarrier beads. This novel approach to metabolic screening allows for the rapid evaluation of the effects of dosing cells with chemical agents.
Schulz
Craig M
CM
University of Washington, Department of Chemistry, PO Box 351700, Seattle, WA 98195, USA.
Scampavia
Louis
L
Ruzicka
Jaromir
J
D000818:Animals; D002460:Cell Line; D002477:Cells / Q000378:metabolism*; D004796:Clinical Enzyme Tests; D003199:Computer Systems; D017022:Flow Injection Analysis / Q000295:instrumentation; D005947:Glucose / Q000032:analysis* / Q000378:metabolism; D022781:Hepatocytes; D019344:Lactic Acid / Q000032:analysis*; D008904:Miniaturization
D019344:Lactic Acid; D005947:Glucose