Large-scale mammalian cell culture: Design and use of an economical batch suspension system

Large-scale mammalian cell culture in the absence of antibiotics requires stringent conditions of sterility for all vessels, procedure, and systems used. Application of existing fermentation technology suffers from the differences between mammalian and bacterial cultures. Relatively simple and inexpensive 100-L vessels have been designed specifically for medium storage and antibiotic-free mammalian cell culture. These vessels are portable and sterilized in a 2 x 3 x 5 ft conventional or VACUMATIC autoclave. They consist of 30-gal 316 stainless-steel sanitary process drums whose heads have been modified to meet the rapid pressure changes that occur during autoclaving. The vessels incorporate systems for aseptic introduction and removal of both liquids and gases required for inoculation, growth, and harvesting of cell suspensions. A two-disk vibromixer is used for agitation with inoculation at a laminar flow hood and incubation in a warm room. These vessels have been used for culture of one rat and eight human tumor lines for over 2 x 10(5) L of suspension.