Flow cytometry and two-dimensional electrophoresis (2-DE) for system evaluation of long term continuous perfused animal cell cultures in macroporous beads.
Immobilization of r-CHO cells at high density using macroporous polyethylene carriers in a modular fluidized bed reactor is demonstrated. Specific growth rates of the cells are measured by incorporation of BrdU. At a cell density of about 10(8) cells/ml a stable growth rate of 0.004 h-1 was established. Total release of proteins into the culture supernatant during protein-free perfusion was analyzed by 2-DE in various phases of the long-term culture showing very similar patterns indicating a constant pattern of gene expression.
Institute of Applied Microbiology (IAM), University of Agriculture, Vienna, Austria.
M
M
Reiter
N
N
Borth
G
G
Blüml
K
K
Wimmer
H
H
Harant
N
N
Zach
T
T
Gaida
C
C
Schmatz
H
H
Katinger
D000818:Animals; D001709:Biotechnology / Q000295:instrumentation*; D016466:CHO Cells / Q000166:cytology*; D002455:Cell Division / Q000502:physiology; D006224:Cricetinae; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D008863:Microspheres; D010477:Perfusion; D016062:Porosity; D013997:Time Factors