Enhancement of Sf-9 cell growth and longevity through supplementation of culture medium with hemolymph.
The benefits of insect cell culture medium supplementation with hemolymph of Lonomia obliqua were investigated. The addition of hemolymph to the medium induced high levels of cell growth, and the viability was maintained for longer periods. The maximum cell yield increased almost 3-fold after hemolymph supplementation. Cultures in their stationary phase were rescued through hemolymph supplementation, also reaching high cell concentrations. These actions were much dependent on the concentration of hemolymph; low hemolymph concentration had a positive effect in cell growth, whereas high hemolymph concentration showed a deleterious effect. Fractionation of hemolymph by gel filtration chromatography showed the presence of three factors with different activity in insect cell culture: an potential anti-apoptotic factor, a growth-promoting factor, and an enzyme that hydrolyzes sucrose. Addition of hemolymph to the medium induced high levels of glucose production. The sucrose to glucose conversion was also linearly dependent upon the hemolymph concentration. Therefore, we conclude that cell growth and longevity can be increased by supplementation of the culture medium with hemolymph.
Maranga
Luis
L
Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica IBET/ITQB, Apartado 12, P-2781-901 Oeiras, Portugal.
Mendonça
Ronaldo Z
RZ
Bengala
André
A
Peixoto
Cristina C
CC
Moraes
Roberto H P
RH
Pereira
Carlos A
CA
Carrondo
Manuel J T
MJ
D000818:Animals; D016367:Baculoviridae / Q000472:pathogenicity; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D005947:Glucose / Q000378:metabolism; D006458:Hemolymph / Q000378:metabolism*; D007915:Lepidoptera / Q000187:drug effects / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development* / Q000821:virology
D017077:Culture Media, Conditioned; D005947:Glucose