High density cultivation of genetically-engineered CHO cell lines with microcarrier culture systems.
Genetically-engineered CHO cell lines, r beta-13 and CLF-8B2, were cultivated with the MC-1 microcarrier culture system. The cell density could be enhanced by increasing the concentration of microcarrier. At a microcarrier concentration of 10 mg/ml, the cell density could reach 4 to 5 x 10(6) cells/ml. It was shown that these cell lines would spontaneously release from the microcarrier to attach to and proliferate on fresh microcarriers. We were thus able to scale up cultivation using a simple method, i.e. by adding fresh microcarriers and medium directly into the culture system to about 2, 4 or 8 times the original volume. Using a perfusion culture system, we have successfully cultivated CLF-8B2 cells in a 2 L bioreactor for several weeks at medium perfusion rates of 0.5 to 3 working volumes. Prourokinase was stably secreted.
Xiao
C
C
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing.
Huang
Z
Z
Liu
F
F
Guo
Z
Z
Gao
L
L
D000818:Animals; D016466:CHO Cells / Q000378:metabolism* / Q000648:ultrastructure; D002455:Cell Division; D006224:Cricetinae; D004792:Enzyme Precursors / Q000096:biosynthesis; D006168:Guinea Pigs; D011994:Recombinant Proteins / Q000096:biosynthesis; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis*
D004792:Enzyme Precursors; D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator; C067551:saruplase