Metabolism of hybridoma cells and antibody secretion at high cell densities in dialysis tubing.
The experimental setup, consisting of a bundle of dialysis tubing 2.5 mm in diameter [10-15 kD cutoff, mean pore size 25 A, 20 microns (dry) and 40 microns (wet) wall thickness] inserted into a 1-l glass bioreactor supplied with oxygen and pH electrodes, a porous gas distributor, a sampling tube, and a holder for the eight pieces of dialysis tubing, was developed to investigate the properties and the microenvironment of hybridoma cells enclosed in the tubing during their batch cultivation. The concentrations of low-molecular-weight medium components were the same inside and outside the tubing, and it was possible to control the microenvironment of the cells in the tubing easily. The cell damage caused by mechanical stress was less in the dialysis tubing than in stirred spinner flasks. The influence of the initial cell density in the range from 4 X 10(5) to 1 X 10(8) cells ml-1 and the cultivation time were evaluated according to the total and viable cell concentrations and the cell/cell fragment size distributions. Furthermore, the cell membrane properties, glucose consumption rate, lactate, ammonia and lipid storage material, and the monoclonal antibody production rates as well as intracellular enzyme activities in the culture medium were measured and compared to those in reference cultures in spinner flasks with the same inoculum at low initial cell densities. In dialysis tubing in a concentration range of 5 X 10(6) to 10(8) cells ml-1, the total and viable concentrations of cells remained the same during cultivation.(ABSTRACT TRUNCATED AT 250 WORDS)
Institut für Technische Chemie, Universität Hannover, FRG.
C
C
Käsehagen
F
F
Linz
G
G
Kretzmer
T
T
Scheper
K
K
Schügerl
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001402:B-Lymphocytes / Q000276:immunology; D002455:Cell Division; D002459:Cell Fusion; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D002470:Cell Survival; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods; D003956:Dialysis; D004553:Electric Conductivity; D006825:Hybridomas / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D010954:Plasmacytoma / Q000276:immunology
D000911:Antibodies, Monoclonal