On-line monitoring of human prostate cancer cells in a perfusion rotating wall vessel by near-infrared spectroscopy.
PC-3 human prostate cancer cells have been cultivated in a rotating wall vessel in which glucose, lactate, and glutamine profiles were monitored noninvasively and in real time by near-infrared (NIR) spectroscopy. The calibration models were based on off-line spectra from tissue culture experiments described previously (Rhiel et al., Biotechnol Bioeng 77:73-82). Monitoring performance was improved by Fourier filtering of the spectra and initial off-set adjustment. The resulting standard errors of predictions were 0.95, 0.74, and 0.39 mM for glucose, lactate, and glutamine, respectively. The concentration of ammonia could not be accurately measured from the same spectra. In addition, metabolite uptake and production rates were determined for PC-3 prostate cancer cells during exponential growth in batch-mode cultivation. Cells grew with a doubling time of 21 h and consumed glucose and glutamine at rates of 6.8 and 1.8 x 10(-17) mol/cell.s, respectively. This resulted in lactate and ammonia production rates of 11.9 and 1.3 x 10(-17) mol/cell.s, respectively. Compared with other monitoring technologies, this technology has many advantages for spaceflights and stand-alone units; for instance, calibration can be performed at one time and then applied in a reagentless, low-maintenance way at a later time. The resulting concentration information can be incorporated into closed-loop control schemes, thereby leading to better in vitro models of in vivo behavior.
Rhiel
Martin H
MH
Department of Chemical and Biochemical Engineering, 4133 Seamens Center, University of Iowa, Iowa City, IA 52242, USA.
Cohen
Michael B
MB
Arnold
Mark A
MA
Murhammer
David W
DW
D000641:Ammonia / Q000032:analysis / Q000378:metabolism; D019149:Bioreactors; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D005973:Glutamine / Q000032:analysis / Q000378:metabolism; D006801:Humans; D019344:Lactic Acid / Q000032:analysis / Q000378:metabolism; D008297:Male; D010477:Perfusion; D011471:Prostatic Neoplasms / Q000378:metabolism*; D019265:Spectroscopy, Near-Infrared
D005973:Glutamine; D019344:Lactic Acid; D000641:Ammonia; D005947:Glucose