Studies of baby hamster kidney natural cell aggregation in suspended batch cultures.
Microcarrier cultures of animal cells of industrial relevance are known to shed aggregates into the suspension phase. For a BHK cell line, which is known to be prone to aggregate naturally, microcarrier and aggregate forms of culture are compared in spinner culture. In microcarrier cultures, it is shown that increasing initial microcarrier concentration yields decreasing concentration of smaller aggregates in suspension; roughly equivalent concentrations of total cells and single cells in suspension are obtained. In the absence of Cytodex 3, aggregate final size is hydrodynamically controlled in batch and semicontinuous suspension culture. Rate of agitation is the main variable controlling aggregate size in batch cultures. The range of agitation rates studied (20 to 70 rpm in 250 mL spinner flasks) produced aggregates with maximum sizes of 200 microns. Necrotic centers were not observed; this was confirmed by Trypan blue viability measurements after mechanical dissociation of aggregates and also by the constant productivity obtained from different aggregate sizes. Comparing aggregate and microcarrier culture conditions, it is shown that at 100 rpm maximum total cell concentration is larger in the absence of microcarriers; dead cell concentrations, most of which exist in suspension, are slightly larger in microcarrier culture. Total viable cell concentrations in aggregate, hydrodynamically controlled culture, are almost one order of magnitude higher than in microcarrier cultures. These results suggest that there might be advantages in using aggregate cultures under hydrodynamic control of aggregate size in lieu of microcarrier cultures for naturally aggregating cell lines.
Instituto de Biologia Experimental Tecnológica, Instituto Tecnologia Química Biológica, Oeiras, Portugal.
J L
JL
Moreira
P M
PM
Alves
J M
JM
Rodrigues
P E
PE
Cruz
J G
JG
Aunins
M J
MJ
Carrondo
D000818:Animals; D002449:Cell Aggregation; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D006224:Cricetinae; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods*; D007668:Kidney / Q000166:cytology*; D008647:Mesocricetus; D010316:Particle Size; D013535:Suspensions
D013535:Suspensions