Submerged Culture of Mammalian Cells: The Five Liter Fermentor,

Abstract

The design and application of a system permitting the active proliferation of mammalian cells, as individual units, in the submerged state, would seem to be of value to individuals interested in investigations relating to the overall kinetics of the host/cell virus interaction and to studies requiring high concentrations of uniform tissue cells or virus. This report presents data establishing the feasibility of obtaining uniform proliferation of mammalian cells in the submerged state in volumes up to 3 L in a stationary, impeller-agitated fermentor. Various types of agitated systems have been suggested as satisfactory for permitting growth of tissue cells in the submerged state, i. e., the rotary shaker of Earle et al. (1954), the tumble tube of Owens et al. (1953), the roller tube of Graham and Siminovitch (1955), the suspended stirrer in Erlenmeyer flasks of Cherry and Hull (1956), Powell's (1954) hexagonal roller tube, Brown and Hardy's (1957) wrist shaker, as well as the glass stirrer of Danes (1957). Inherent difficulties have been encountered with certain of the submerged culture systems mentioned. Thus, in shake flask systems there is a marked tendency toward the formation of large granules (McLimans et al., 1957). Additionally, proteinaceous precipitates are frequently formed. Certainly, the extrapolation to large vessels, 3 to 16 L, would be difficult in the instance of roller-tube type units or flask systems. We have reported (McLimans et al., 1957), l Contribution No. 12 from Microbiology in Medicine, Wistar. The investigations herein reported were conducted under the auspices of a grant (Contract no. DA-18-064-404-CML-74) from the U. S. Army Chemical Corps, Fort Detrick, Maryland.

Effect of ouabain and metabolic inhibitors on the Na and K movements of nucleotide contents of L cells

“…LS cells. Cells were grown in stirrer cultures in a 1 1. aspirator bottle stirred with a Teflon magnet with an inlet and outlet for gassing and an inlet and outlet for feeding and withdrawing the cells (see Danes, 1957;McLimus, Davis, Glover & Rake, 1957). The medium used was Eagle's basal medium (BME) plus glutamine (1.6 mm), and calf serum 5 %, obtained from Flow Laboratories.…”

Effects of Tannic Acid and Its Related Compounds upon Chikungunya Virus

“…The purpose of our present study, therefore, was to examine further the mechanisms of virus-inactivating actions of tannins, by dealing with the interaction of Chikun-gunya virus (a member of Alphavirus) with tannic acid (belonging to hydrolyzable tannins) and its related compounds. MATERIALS AND METHODS (17).…”

Breast milk cell supernatants from atopic donors stimulate cord blood IgE secretion in vitro

“…Cord blood lymphocytes were isolated from Ficoll/Hypaque density gradient sedimentation (Boyum, 1968) from 30 ml of heparinized (preservative-free) blood drawn from nine separate placentas. Cell identification and counting were performed as before and viability was assessed by trypan-blue exclusion (McLimans et al, 1957). The atopie status of each of the CBL donor's parents was assessed by interview as well as medical records whenever possible.…”