Preservation of Mammalian Cells in a Chemically Defined Medium and Dimethylsulfoxide

Brown, Bruce L.; Nagle, Stanley C.

doi:10.1126/science.149.3689.1266

Cited by 18 publications

(5 citation statements)

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“…DMSO is the most commonly used cryoprotectant for storing cells [Brown and Nagle, ; Fahmy et al, ]. The TGF‐β enhancer activity of DMSO toward different cell types [Brown and Nagle, ; Fahmy et al, ] would typically increase survival of these cells during the freezing and thawing processes. The solution commonly used for freezing cells consists of 30% fetal calf serum which contains TGF‐β mainly derived from plasma platelets.…”

Section: Resultsmentioning

confidence: 99%

“…DMSO has been used as a cryoprotectant when added to cell media. It reduces intracellular ice formation and thus prevents cell death during the freezing process [Brown and Nagle, ]. To see if other cryoprotectants exhibit the same TGF‐β enhancer activity, we determined the effects of 1% (v/v) of DMSO, glycerol, ethylene glycol, propylene glycol, and formamide [Fahmy et al, ] on TGF‐β‐stimulated luciferase activity in MLE cells.…”

Section: Resultsmentioning

confidence: 99%

“…Among these agents, only DMSO enhanced TGF-b-stimulated luciferase activity (data not shown). DMSO is the most commonly used cryoprotectant for storing cells [Brown and Nagle, 1965;Fahmy et al, 2014]. The TGF-b enhancer activity of DMSO toward different cell types [Brown Fig.…”

Section: Journal Of Cellular Biochemistrymentioning

confidence: 99%

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DMSO Enhances TGF‐β Activity by Recruiting the Type II TGF‐β Receptor From Intracellular Vesicles to the Plasma Membrane

Huang

Chen

Huang

et al. 2016

J of Cellular Biochemistry

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Dimethyl sulfoxide (DMSO) is used to treat many diseases/symptoms. The molecular basis of the pharmacological actions of DMSO has been unclear. We hypothesized that DMSO exerts some of these actions by enhancing TGF-β activity. Here we show that DMSO enhances TGF-β activity by ~3–4-fold in Mv1Lu and NMuMG cells expressing Smad-dependent luciferase reporters. In Mv1Lu cells, DMSO enhances TGF-β-stimulated expression of P-Smad2 and PAI-1. It increases cell-surface expression of TGF-β receptors (TβR-I and/or TβR-II) by ~3–4-fold without altering their cellular levels as determined by 125I-labeled TGF-β-cross-linking/Western blot analysis, suggesting the presence of large intracellular pools in these cells. Sucrose density gradient ultracentrifugation/Western blot analysis reveals that DMSO induces recruitment of TβR-II (but not TβR-I) from its intracellular pool to plasma-membrane microdomains. It induces more recruitment of TβR-II to non-lipid raft microdomains than to lipid rafts/caveolae. Mv1Lu cells transiently transfected with TβR-II-HA plasmid were treated with DMSO and analyzed by indirect immunofluoresence staining using anti-HA antibody. In these cells, TβR-II-HA is present as a vesicle-like network in the cytoplasm as well as in the plasma membrane. DMSO causes depletion of TβR-II-HA-containing vesicles from the cytoplasm and co-localization of TβR-II-HA and cveolin-1 at the plasma membrane. These results suggest that DMSO, a fusogenic substance, enhances TGF-β activity presumably by inducing fusion of cytoplasmic vesicles (containing TβR-II) and the plasma membrane, resulting in increased localization of TβR-II to non-lipid raft microdomains where canonical signaling occurs. Fusogenic activity of DMSO may play a pivotal role in its pharmacological actions involving membrane proteins with large cytoplasmic pools.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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DMSO Enhances TGF‐β Activity by Recruiting the Type II TGF‐β Receptor From Intracellular Vesicles to the Plasma Membrane

Huang

Chen

Huang

et al. 2016

J of Cellular Biochemistry

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“…This loss in virus yield is due to initial freezing along with cellular damage from decay of 32P that has been incorporated by the cells. The unincorporated 32P in these cultures amounts to 5 pcuries/ml and this level of radioactivity has no effect upon the capacity of cells to 3. Viral growth in 32P-labeled cells.…”

Section: Labeling Of Early Viral Rnamentioning

confidence: 98%

“…Dimethylsulfoxide (DMSO) has been shown to serve as an excellent preservative for the freezing of animal cells (3,9,11,17); therefore experiments were performed with DMSO to determine whether HeLa S3 cells could be frozen and thawed without extensive loss of viability. Four methods were used to measure cell viability: (1) the exclusion of erythrosin B dye, (2) plating efficiencies, (3) infectious center titrations after infection with poliovirus and (4) virus yield.…”

Section: Measurement Of Cell Viability After Freezingmentioning

confidence: 99%

Stabilization of poliovirus to ³²P decay

Hughes¹,

Tershak²

1971

Can. J. Microbiol.

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1971. Stabilization of poliovirus to 32P decay. Can. J. . Suicide experiments with 32P-labeled poliovirus were performed with HeLa S3 cells. The results show that virus-cell complexes become increasingly refractory to 32P decay during the eclipse period, i.e., stabilization to 32P decay occurs during the first 3 h of infection. The data suggest that stabilization is probably due in part to the production of early RNA during the eclipse period because the presence of 32P in the medium during the first 2 h of growth sensitizes infective centers. IntroductionIn 1947 Luria and Latarjet (10) designed an experiment to detect an increase in the number of intracellular phage particles by subjecting phagehost complexes to increasing doses of ultraviolet (uv.) irradiation after infection. Since that time experiments have been performed with uv. light, high-energy X-rays, and internal radiation resulting from the disintegration of incorporated 32P atoms. The data have shown that as intracellular phage development proceeds, a progressive resistance to irradiation occurs (2, 10, 12, 13, 14, 15).To our knowledge no progressive stabilization to 32P decay has been reported with cells infected with an animal virus. Henry and Youngner (7) performed studies with HeLa cells and 32P-labeled poliovirus RNA, but an experiment comparable to that with bacteriophage was not possible because mammalian cells are sensitive to freezing. Therefore modifications were innovated and a temperature of 2C was chosen to arrest viral development. The results failed to show an appreciable increase in resistance to 32P decay during the eclipse period. Experiments with both control and 32P-labeled RNA showed that intracellular viral RNA is unstable at 2C for long periods of time, and the authors suggested that enzymatic degradation might play a part in the sensitivity of intracellular RNA at this temperature.With the use of dimethylsulfoxide (DMSO) a freezing process has been developed which permits poliovirus infected cells to be frozen and

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Dimethyl Sulfoxide in Inorganic Chemistry

Reynolds

1970

Progress in Inorganic Chemistry

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Preservation of Mammalian Cells in a Chemically Defined Medium and Dimethylsulfoxide

Cited by 18 publications

References 1 publication

DMSO Enhances TGF‐β Activity by Recruiting the Type II TGF‐β Receptor From Intracellular Vesicles to the Plasma Membrane

DMSO Enhances TGF‐β Activity by Recruiting the Type II TGF‐β Receptor From Intracellular Vesicles to the Plasma Membrane

Stabilization of poliovirus to ³²P decay

Dimethyl Sulfoxide in Inorganic Chemistry

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