Production of Mouse Interferon with High Titers in a Large‐Scale Suspension Culture System1

Sano, Emiko; Matsui, Yoshiko; Kobayashi, S

doi:10.1111/j.1348-0421.1974.tb00805.x

Cited by 9 publications

(7 citation statements)

References 18 publications

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“…Mouse L, L-MS [11], rabbit It has been reported that pretreatment with IF caused an earlier IF production [6,10,13]. We also examined the kinetics of IF production in L cells pretreated or untreated with IF (Fig.…”

Section: Cell Culturesmentioning

confidence: 99%

Enhancing Effect of Interferon Pretreatment on Interferon Production

Ito

Kobayashi

1974

Japanese Journal of Microbiology

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Section: Cell Culturesmentioning

confidence: 99%

Enhancing Effect of Interferon Pretreatment on Interferon Production

Ito

Kobayashi

1974

Japanese Journal of Microbiology

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“…The uppermost tray is connected with inlet/outlet parts (6) to its vertical holes. One of the parts is connected to reservoir (7) with silicone rubber tubing (8), and the other to air filter (9). layer on each tray after the incubation for 5-6 days at 37CC.…”

Section: Methodsmentioning

confidence: 99%

Interferon Production with Multitray Culture System on a Large Scale

Utsumi¹,

Iizuka²,

Kobayashi³

1984

Journal of Interferon Research

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A multitray culture system was devised for cell culture and interferon (IFN) production on a large scale. The system consisted of a cell culture apparatus which has stacked multitrays and a reservoir. All components of the system were made of nontoxic, nonmutagenic, washable, and autoclavable materials, so that the system can be repetitively used in suitable condition for cultivations of anchorage-dependent cells with low running cost. Large quantities of mouse IFN (containing both MuIFN-alpha and MuIFN-beta) from mouse L cells and human beta interferon (HuIFN-beta) from human fibroblast cells, respectively, were obtained very efficiently using this system in comparison with roller bottle, bulk culture vessel, and microcarrier systems.

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“…Mouse L cells, human FL cells and rabbit RK-13 cells were employed for assaying IF preparations of the corresponding origin. Two different lines of mouse L cells (L-MS cells) and human FL cells (FL-MS cells) had been adapted to grow readily either in monolayer or in suspension in our laboratory [9]. The growth medium employed for monolayer culture was Eagle's minimum essential medium (Nissui Seiyaku Co., Japan) plus 5% calf serum (MEM + 5% CS).…”

Section: Methodsmentioning

confidence: 99%

“…Virus. Vesicular stomatitis virus (VSV), New Jersey serotype, propagated in a human embryo lung cell line (HELK cells, obtained from Dr. Yoshioka of the Kitasato Institute, Tokyo, Japan) was used as the challenge virus in the IF assay [9].…”

Section: Methodsmentioning

confidence: 99%

A Rapid and Simple Method for Assaying Interferon1

Suzuki

Akaboshi

Kobayashi

1974

Japanese Journal of Microbiology

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A new procedure of assaying interferon (IF) has been developed. Cell suspension was dispensed into liquid scintillation counting vials together with IF sample. During an overnight incubation, the cells adhered sufficiently to the bottom of the vials and all the subsequent procedures were carried out without transfer of the cells from the vials. Vesicular stomatitis virus was inoculated and virusspecific RNA was labeled by adding 3H-uridine and actinomycin D to the medium. Incubation was terminated prior to completion of a single-step growth of the virus and radioactivity of the labeled cells in each vial was determined.The reciprocal of the IF dilution which reduced the radioactivity in viral RNA by 50% was taken as the titer. The present procedure consists of simple manipulations and can be completed within 24 hr. Furthermore, it is quite reproducible and gives a titer almost identical to that obtained by the conventional plaque-reduction dose method. The procedure can be applied to mouse L cells, rabbit RK-13 cells and human FL cells, without modification.The technique which is currently used most widely for assaying interferon (IF) is the plaque-reduction dose method (PRD50 method). The procedure gives reliable results but involves a longer time and laborious handlings. In order to overcome these disadvantages, several groups of workers have introduced techniques based on inhibition of nucleic acid synthesis (INAS50 method) [1,4 6], in which the IF titer is determined by measuring radioactivity of 3H-uridine incorporated into virus-specific RNA in the IFtreated cells. Although their methods have some advantages over the PRD50 method, these are still laborious and remain impractical as a routine method. In this communication we describe a new procedure of the INAS50 method which yields reliable results much faster in a simpler way. This technique will be well qualified to replace the PRD50 method. MATERIALS AND METHODSCells and media. Mouse L cells, human FL cells and rabbit RK-13 cells were employed for assaying IF preparations of the corresponding origin. Two different lines of mouse L cells (L-MS cells) and human FL cells (FL-MS cells) had been adapted to grow readily either in monolayer or in suspension in our laboratory [9]. The growth medium employed for monolayer culture was Eagle's minimum essential medium (Nissui Seiyaku Co., Japan) plus 5% calf serum (MEM + 5% CS). For suspension culture Eagle's spinner culture medium (Nissui Seiyaku Co., Japan) supplemented with 5% calf serum (SM+5%CS) was used.

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Production of Mouse Interferon with High Titers in a Large‐Scale Suspension Culture System1

Cited by 9 publications

References 18 publications

Enhancing Effect of Interferon Pretreatment on Interferon Production

Enhancing Effect of Interferon Pretreatment on Interferon Production

Interferon Production with Multitray Culture System on a Large Scale

A Rapid and Simple Method for Assaying Interferon1

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