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Merck

Novel in vitro platform to investigate myotube atrophy.

Anticancer research (2015-04-12)
Christopher Oelkrug, Katharina Horn, Gustavo R Makert, Andreas Schubert
RESUMEN

The electrical current exclusion (ECE) principle provides an alternative to common methods of cell diameter measurement and especially in atrophy and cancer associated cachexia research. C2C12 myoblasts were differentiated into myotubes and treated with 100 μM dexamethasone to induce atrophy in vitro. Subsequently, they were incubated for 24 h with media containing different concentrations of curcumin and/or branched-chain amino acids (BCAAs) in order to counteract atrophy. After treatment with curcumin, an increase in cell diameter was detectable; the highest increase with 13.9 ± 0.4% was seen with 10 μM curcumin. The combination of curcumin and BCAAs showed an increase of 13.4 ± 1.2 %. Cell diameter measurement via the ECE showed that curcumin, and curcumin in combination with BCAAs, were able to restore atrophic C2C12 myotubes. Therefore, the application of ECE in muscle atrophy and also cancer-associated cachexia research allows rapid screening of novel compounds in order to test their efficacy in vitro.

MATERIALES
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Sigma-Aldrich
Dodecilsulfatosódico, BioReagent, suitable for electrophoresis, Molecular Biology, ≥98.5% (GC)
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Clorhidrato de Trizma®, reagent grade, ≥99.0% (titration), crystalline
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Dexametasona, powder, BioReagent, suitable for cell culture, ≥97%
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Dodecilsulfatosódico, ≥99.0% (GC), dust-free pellets
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Cloruro de sodio, Molecular Biology, DNase, RNase, and protease, none detected, ≥99% (titration)
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Sodium chloride solution, 5 M in H2O, BioReagent, Molecular Biology, suitable for cell culture
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Cloruro de sodio, BioXtra, ≥99.5% (AT)
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Cloruro de sodio, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
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Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture
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Dodecilsulfatosódico, ACS reagent, ≥99.0%
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Sodium dodecyl sulfate solution, BioUltra, Molecular Biology, 10% in H2O
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Clorhidrato de Trizma®, BioPerformance Certified, suitable for cell culture, ≥99.0% (titration)
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Dodecilsulfatosódico, ReagentPlus®, ≥98.5% (GC)
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L-Leucine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
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Curcumin, from Curcuma longa (Turmeric), powder
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L-Isoleucina, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
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Sodium chloride solution, 5 M
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Selenium, powder, −100 mesh, 99.99% trace metals basis
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Dexametasona, ≥98% (HPLC), powder
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Dodecilsulfatosódico, BioUltra, Molecular Biology, ≥99.0% (GC)
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Sodium dodecyl sulfate solution, BioUltra, 20% in H2O
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L-Leucine, reagent grade, ≥98% (HPLC)
SAFC
Sodium chloride solution, 5 M
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L-Valine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
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Dodecilsulfatosódico, dust-free pellets, suitable for electrophoresis, Molecular Biology, ≥99.0% (GC)
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Selenium, powder, −100 mesh, ≥99.5% trace metals basis
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Dodecilsulfatosódico, suitable for ion pair chromatography, LiChropur, ≥99.0%
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Cloruro de sodio, BioUltra, Molecular Biology, ≥99.5% (AT)
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Curcumin, ≥94% (curcuminoid content), ≥80% (Curcumin)
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Cloruro de sodio, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%