[Effects of hydrodynamic on aggregates formation, growth and metabolism of HEK293 cells in suspension culture].
By using the size distribution of cell aggregates, viable cell density, cell viability, specific consumption rate of glucose (q(glc)), specific production rate of lactate (q(lac)) and lactate transform rate (Y(lac/glc)) as the evaluation indexes, the effects of hydrodynamic on aggregates formation, growth and metabolism of HEK293 cells in suspension culture were examined in 250mL spinner-flasks by setting the agitation rates at 25, 50, 75 and 100r/min, respectively. It was found that agitation plays an important role in HEK293 cell aggregates formation and cell aggregates size distribution. After 7d cultivation in spinner-flasks operated at 50r/min and 75r/min, the average diameter of HEK293 cell aggregates was 201 microm and 175 microm, respectively, with the fraction of aggregates larger than 225 microm less than 10%. The cell viability was kept above 90% with the metabolic indexes, including q(glc), q(lac) and Y(lac/glc) kept constant. These results demonstrated that hydrodynamic derived from the proper agitation play a decisive role in controlling the formation and size distribution of HEK293 cell aggregates, and provided sufficient mass transfer to support the normal growth and metabolism of HEK293 cells in suspended aggregates.
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 10071, China.
Hong
H
Liu
Xing-Mao
XM
Liu
Ben-Chuan
BC
Wu
Ling-Ling
LL
Ye
Xiao-Ping
XP
Ni
Pei-Tang
PT
Huang
Zhao-Lie
ZL
Chen
D019149:Bioreactors; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000295:instrumentation; D002460:Cell Line; D049109:Cell Proliferation; D006801:Humans; D007700:Kinetics