Growth ofChlamydia psittaci strain menigopneumonitis in mouse L cells cultivated in a defined medium in spinner cultures

Morrison, Susan J.; Jenkin, Howard M.

doi:10.1007/bf02615966

Cited by 30 publications

(6 citation statements)

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“…Our results extended the use of L cells grown in a serum-free medium in a spinner culture (17) to their successful use in a shaker culture (10). With this culture system we were able to obtain equal or higher titers of C. trachomatis LGV-404L than those observed with other methods of cultivation, as well as organisms free of serum contamination and adventitious agents from serum.…”

Section: Discussionmentioning

confidence: 54%

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Use of cycloheximide to study independent lipid metabolism of Chlamydia trachomatis cultivated in mouse L cells grown in serum-free medium

Reed¹,

Anderson²,

Jenkin³

1981

Infect Immun

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A system for measuring chlamydial lipid synthesis was developed with mouse L cells grown in serum-free modified Waymouth 752/l medium in a shaker culture. Host lipid synthesis was reduced approximately 90% when cells were incubated for 24 h in medium containing cycloheximide (2 micrograms/ml). Lipid metabolism was monitored by measuring the incorporation of [3H]isoleucine into the total lipid of normal and infected cells. The results suggested that lipid synthesis of Chlamydia trachomatis lymphogranuloma venereum (LGV-404L) was not inhibited by cycloheximide treatment when the chlamydiae were grown in L cells, whereas host lipid synthesis was inhibited. Chlamydial lipid metabolism began about 6 to 12 h after infection when the noninfectious reticulate body was found and continually increased until the beginning of the appearance of intracellular infectious elementary bodies at 24 to 30 h.

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Section: Discussionmentioning

confidence: 54%

“…Cx-treated L cells grown in modified Waymouth 752/1 (WO5) medium (17) without serum or undefined exogenous lipids provides an ideal system for studying chlamydial lipid synthesis. The shaker culture system provides a means of obtaining high yields of cells and chlamydiae with a minimal chance of contamination (10).…”

mentioning

confidence: 99%

Use of cycloheximide to study independent lipid metabolism of Chlamydia trachomatis cultivated in mouse L cells grown in serum-free medium

Reed¹,

Anderson²,

Jenkin³

1981

Infect Immun

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“…Culture medium A modified HI/W05/BA2ooo (Morrison and Jenkin 1972) without serum or hormones was employed throughout the experiments as the basal culture medium. HI/WO5/BA2ooo was prepared by adding bovine serum albumin (Pentex, Fraction V, fatty acid free, Miles Laboratories, Elkhart, Ind., 200 mg/1) and oleic acid (Na salt, Sigma Chem.…”

Section: Preparation Of Insulin-dextran Complexesmentioning

confidence: 99%

The effect of insulin-dextran complexes on the protein synthesis in the primary monolayer culture of adult rat hepatocytes.

Nakai

Kutsumi

Sakamoto

et al. 1984

Tohoku J. Exp. Med.

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The purpose of the present study was to investigate the mechanisms of the long-term effects of insulin on protein synthesis in rat liver parenchymal cells. Three kinds of soluble and stable insulin-dextran complexes (I: Mr 150, 000; II: Mr 450,000; III : Mr 2,000,000) were prepared. The effects of insulin or insulin-dextran complexes on protein synthesis were evaluated in the primary monolayer culture of adult rat liver. To avoid the effects of serum factors, the primary monolayer culture of adult rat liver in serum free medium was established. The cultured hepatocytes well maintained the metabolic and morphological characteristics of the adult rat liver. The incorporation of ['SC] -leucine into trichloroacetic acid insoluble proteins and immunoprecipitates by anti-rat albumin serum were measured in cultured hepataocytes prepared either from the control or streptozotocin-induced rats. An addition of insulin-dextran complexes to the culture medium of the hepatocytes from diabetic rats stimulated the protein synthesis as well as the native insulin. The maximal effect of insulin-dextran complex (I) on protein synthesis was comparable to native insulin. However, insulin-dextran complex (II) caused a 67% stimulation of native insulin. Insulindextran complexes (III) induced only a slight increase of protein synthesis. Furthermore, an addition of insulin-dextran complexes into the medium caused a more tight cellular attachment to the dish and more extensive spreading. These results favor the view that the stimulation by insulin of protein synthesis in rat hepatocytes does not require the entry of insulin into cells, insulin ; protein synthesis ; cultured hepatocytes

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“…Thus, there would be advantages in removing calf serum, or all unnecessary components of serum, from tissue culture media when studying the biological characteristics of chlamydiae growing within mammalian cells. For example, Morrison & Jenkin ( 1972) found that the lipid metabolism of C. psittaci in suspended L-cell cultures could not be assessed in serum-containing media. In their short-term metabolic experiments serum could be replaced by bovine serum albumin plus sodium oleate.…”

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confidence: 99%

The Role of Calf Serum in the Growth of Chlamydia trachomatis in McCoy Cell Cultures

Karayiannis

Hobson

1981

Microbiology

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Chlamydia trachomatis is normally grown in McCoy monolayer cover slip cultures using partially defined media containing foetal calf serum at concentrations up to 10% (v/v). Omission of the serum decreased the number of inclusions produced by infecting the McCoy cells with a standard inoculum of a genital strain of C. trachomatis. Substitution of the foetal calf serum with a macromolecular fraction from the serum or with a mixture of sodium oleate, bovine serum albumin fraction V and fetuin maintained the inclusion count, but substitution with serum filtrate, containing the amino acids and other low molecular weight components, decreased the inclusion count of C. trachomatis. The role of calf serum and the need for a fully defined medium excluding serum for studying the growth of C. trachomatis in tissue culture are discussed.

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Growth ofChlamydia psittaci strain menigopneumonitis in mouse L cells cultivated in a defined medium in spinner cultures

Cited by 30 publications

References 30 publications

Use of cycloheximide to study independent lipid metabolism of Chlamydia trachomatis cultivated in mouse L cells grown in serum-free medium

Use of cycloheximide to study independent lipid metabolism of Chlamydia trachomatis cultivated in mouse L cells grown in serum-free medium

The effect of insulin-dextran complexes on the protein synthesis in the primary monolayer culture of adult rat hepatocytes.

The Role of Calf Serum in the Growth of Chlamydia trachomatis in McCoy Cell Cultures

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