On-line determination of animal cell concentration.

A new approach for the indirect determination of cell concentration in the case of nonconstant metabolic rates has been developed. The specific glucose-uptake rate was shown to be nonconstant in batch cultures of free suspended and immobilized CHO SSF3 cells. Time-independent models correlating the specific rate to the limiting substrate concentration were established, thus providing a continuous determination of the specific rate through on-line measurement of the limiting substrate. The method could be applied to determine on-line cell concentration in both free suspended and immobilized cell cultures. Results were verified off-line by crystal violet nuclei counting. The predicted cell concentration was in very good agreement with the off-line reference during the whole exponential-growth phase, until the specific glucose-uptake rate tended to zero.

[1]  T. Mosmann Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[2]  E. Nicolau,et al.  Growth models , 1987 .

[3]  B C Blake-Coleman,et al.  On‐line monitoring of cell mass in mammalian cell cultures by acoustic densitometry , 1989, Biotechnology and bioengineering.

[4]  R. Margis,et al.  Quantification of attached cells in tissue culture plates and on microcarriers. , 1989, Analytical biochemistry.

[5]  C. Demangel,et al.  Comparison of various methods for monitoring hybridoma cell proliferation. , 1990, Journal of immunological methods.

[6]  K. K. Frame,et al.  Cell volume measurement as an estimation of mammalian cell biomass , 1990, Biotechnology and bioengineering.

[7]  Christopher L. Davey,et al.  Real-time monitoring of cellular biomass: Methods and applications , 1990 .

[8]  K. K. Frame,et al.  Kinetic study of hybridoma cell growth in continuous culture. I. A model for non‐producing cells , 1991, Biotechnology and bioengineering.

[9]  C. Demangel,et al.  Lactate dehydrogenase (LDH) activity of the number of dead cells in the medium of cultured eukaryotic cells as marker , 1992 .

[10]  W. S. Hu,et al.  Cultivation of mammalian cells on macroporous microcarriers. , 1992, Enzyme and microbial technology.

[11]  G Locher,et al.  On-line measurement in biotechnology: techniques. , 1992, Journal of biotechnology.

[12]  K B Konstantinov,et al.  On‐line monitoring of hybridoma cell growth using a laser turbidity sensor , 1992, Biotechnology and bioengineering.

[13]  B Schulze,et al.  On-line immunoanalysis for bioprocess control. , 1993, Journal of biotechnology.

[14]  P. Nabet,et al.  Search for cell proliferation markers suitable for cell count in continuous immobilized cell bioreactors. , 1993, Journal of biotechnology.

[15]  J. Ljunggren,et al.  Catabolic control of hybridoma cells by glucose and glutamine limited fed batch cultures , 1994, Biotechnology and bioengineering.

[16]  R. Pörtner,et al.  Estimation of specific glucose uptake rates in cultures of hybridoma cells. , 1994, Journal of biotechnology.

[17]  B Becker,et al.  In situ screening assay for cell viability using a dimeric cyanine nucleic acid stain. , 1994, Analytical biochemistry.

[18]  K Konstantinov,et al.  Real-time biomass-concentration monitoring in animal-cell cultures. , 1994, Trends in biotechnology.

[19]  Thibaud Stoll Process development studies for the production of monoclonal mucosal IgA antibodies , 1995 .

[20]  A. Zeng,et al.  Mathematical modeling and analysis of glucose and glutamine utilization and regulation in cultures of continuous mammalian cells , 1995, Biotechnology and bioengineering.

[21]  Bernd Hitzmann,et al.  The automation of immun-FIA-systems , 1995 .

[22]  H. McCormack,et al.  A rapid in situ, colorimetric assay for the determination of mammalian cell viability in alginate-immobilized and encapsulated systems , 1996 .

[23]  M. Butler,et al.  Cell Productivity is Overestimated by the Crystal Violet Nuclei Counting Technique , 1997 .

[24]  J. Nielsen,et al.  On-line and in situ monitoring of biomass in submerged cultivations , 1997 .

[25]  U von Stockar,et al.  Measurement of volumetric (OUR) and determination of specific (qO2) oxygen uptake rates in animal cell cultures. , 1998, Journal of biotechnology.

[26]  I. Marison,et al.  High-sensitive heat-flow calorimetry , 1998 .

[27]  A. Zeng,et al.  Variation of Stoichiometric Ratios and Their Correlation for Monitoring and Control of Animal Cell Cultures , 1998, Biotechnology progress.

[28]  R. Kemp,et al.  Specific heat flow rate: an on-line monitor and potential control variable of specific metabolic rate in animal cell culture that combines microcalorimetry with dielectric spectroscopy. , 1998, Biotechnology and bioengineering.

[29]  I. Marison,et al.  A new method for on-line measurement of the volumetric oxygen uptake rate in membrane aerated animal cell cultures. , 2000, Journal of biotechnology.