Quantitative on-line monitoring of cellular glucose and lactate metabolism in vitro with slow perfusion.
An on-line in vitro perfusion technique is described that allows the continuous quantification of cellular glucose metabolism in vitro. Using biosensor technology, we measure glucose and lactate metabolism at a minute-to-minute time resolution for periods up to several days. The application of our perfusion-detection technique for in vitro monitoring is demonstrated in a wide variety of cells, including primary neuronal and astroglia cultures, yeast cells, and human lymphocytes. The method shows that variations in oxygen delivery or exposure to a noncompetitive pseudosubstrate (here 2-deoxyglucose) affects normal glucose metabolism. The innovative advantage of the present system is that, in contrast to other devices including a recently described system, metabolism per cell can be quantified. The potential of in vitro on-line monitoring is discussed for application in studying normal and abnormal metabolism, toxic and nontoxic drug effects, and human tissue biopsies.
Leegsma-Vogt
Gea
G
Department of Psychiatry, Section Biological Psychiatry, University of Groningen, Groningen, The Netherlands.
Venema
Kor
K
Brouwer
Nieske
N
Gramsbergen
Jan Bert
JB
Copray
Sjef
S
Korf
Jakob
J
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D015374:Biosensing Techniques; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002478:Cells, Cultured; D004734:Energy Metabolism; D005947:Glucose / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry / Q000378:metabolism*; D008214:Lymphocytes / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010477:Perfusion; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D019344:Lactic Acid; D005947:Glucose