Effects of temperature shift on cell cycle, apoptosis and nucleotide pools in CHO cell batch cultures

被引:138
作者
Moore, A
Mercer, J
Dutina, G
Donahue, CJ
Bauer, KD
Mather, JP
Etcheverry, T
Ryll, T
机构
[1] GENENTECH INC,DEPT CELL BIOL,S SAN FRANCISCO,CA 94080
[2] GENENTECH INC,DEPT IMMUNOL,S SAN FRANCISCO,CA 94080
关键词
apoptosis; programmed cell death; nucleotides; energy charge; CHO cells; batch culture;
D O I
10.1023/A:1007919921991
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Temperature reduction in CHO cell batch culture may be beneficial in the production of recombinant protein and in maintenance of viability. The effects on cell cycle, apoptosis and nucleotide pools were studied in cultures initiated at 37 degrees C and temperature shifted to 30 degrees C after 48 hours. In control cultures maintained at 37 degrees C, viable cells continued to proliferate until the termination of the culture, however, temperature reduction caused a rapid decrease in the percent of cells in S phase and accumulation of cells in G(1). This was accompanied by a concurrent reduction in U ratio (UTO/UDP-GNAc), previously shown to be a sensitive indicator of growth rate. Culture viability was extended following temperature shift, as a result of delayed onset of apoptosis, however, once initiated, the rate and manner of cell death was similar to that observed at 37 degrees C. All nucleotide pools were similarly degraded at the time of apoptotic cell death. Temperature reduction to 30 degrees C did not decrease the energy charge of the cells, however, the overall rate of metabolism was reduced. The latter may be sufficient to extend culture viability via a reduction in toxic metabolites and/or limitation of nutrient deprivation. However, the possibility remains that the benefits of temperature reduction in terms of both viability and productivity are more directly associated with cultures spending extended time in G(1).
引用
收藏
页码:47 / 54
页数:8
相关论文
共 30 条
[1]   EFFECT OF TEMPERATURE ON NUCLEOTIDE POOLS AND MONOCLONAL-ANTIBODY PRODUCTION IN A MOUSE HYBRIDOMA [J].
BARNABE, N ;
BUTLER, M .
BIOTECHNOLOGY AND BIOENGINEERING, 1994, 44 (10) :1235-1245
[2]   EFFECT OF TEMPERATURE ON HYBRIDOMA CELL-CYCLE AND MAB PRODUCTION [J].
BLOEMKOLK, JW ;
GRAY, MR ;
MERCHANT, F ;
MOSMANN, TR .
BIOTECHNOLOGY AND BIOENGINEERING, 1992, 40 (03) :427-431
[3]   THE POTENTIAL OF FLOW CYTOMETRIC ANALYSIS FOR THE CHARACTERIZATION OF HYBRIDOMA CELLS IN SUSPENSION-CULTURES [J].
COCOMARTIN, JM ;
OBERINK, JW ;
VANDERVELDENDEGROOT, TAM ;
BEUVERY, EC .
CYTOTECHNOLOGY, 1992, 8 (01) :65-74
[4]   MATHEMATICAL-ANALYSIS OF DNA DISTRIBUTIONS DERIVED FROM FLOW MICROFLUOROMETRY [J].
DEAN, PN ;
JETT, JH .
JOURNAL OF CELL BIOLOGY, 1974, 60 (02) :523-527
[5]   LONG-TERM PERFUSION CULTURE OF HYBRIDOMA - A GROW OR DIE CELL-CYCLE SYSTEM [J].
DELABROISE, D ;
NOISEUX, M ;
LEMIEUX, R ;
MASSIE, B .
BIOTECHNOLOGY AND BIOENGINEERING, 1991, 38 (07) :781-787
[6]   FRAGMENTED DNA AND APOPTOTIC BODIES DOCUMENT THE PROGRAMMED WAY OF CELL-DEATH IN HYBRIDOMA CULTURES [J].
FRANEK, F ;
VOMASTEK, T ;
DOLNIKOVA, J .
CYTOTECHNOLOGY, 1992, 9 (1-3) :117-123
[7]  
GIARD DJ, 1982, P SOC EXP BIOL MED, V170, P155
[8]   DETERMINATION OF CELL-LYSIS AND DEATH KINETICS IN CONTINUOUS HYBRIDOMA CULTURES FROM THE MEASUREMENT OF LACTATE-DEHYDROGENASE RELEASE [J].
GOERGEN, JL ;
MARC, A ;
ENGASSER, JM .
CYTOTECHNOLOGY, 1993, 11 (03) :189-195
[9]  
GORCZYCA W, 1993, LEUKEMIA, V7, P659
[10]  
GORZYCA W, 1992, INT J ONCOL, V1, P639