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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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Supelco
Curcumin, suitable for matrix substance for MALDI-MS, ≥99.5% (HPLC)
USP
Dexamethasone, United States Pharmacopeia (USP) Reference Standard
Dexamethasone, European Pharmacopoeia (EP) Reference Standard
USP
Curcumin, United States Pharmacopeia (USP) Reference Standard
Sodium laurilsulfate, European Pharmacopoeia (EP) Reference Standard
Dexamethasone, British Pharmacopoeia (BP) Assay Standard
Curcumin, primary reference standard
Supelco
Dexamethasone, Pharmaceutical Secondary Standard; Certified Reference Material
Dexamethasone for system suitability, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
L-Isoleucine, 99%, FCC, FG
Supelco
Curcumin, analytical standard
Sigma-Aldrich
L-Isoleucine, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, 20% in H2O
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, Molecular Biology, 10% in H2O
Sigma-Aldrich
Curcumin, from Curcuma longa (Turmeric), powder
Sigma-Aldrich
L-Isoleucine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%
Supelco
Sodium chloride, Pharmaceutical Secondary Standard; Certified Reference Material
Isoleucine, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Selenium, pellets, <5 mm particle size, ≥99.999% trace metals basis
Sigma-Aldrich
L-Leucine, 99%, FG
Sigma-Aldrich
Sodium dodecyl sulfate, ACS reagent, ≥99.0%
Sigma-Aldrich
Trizma® hydrochloride, reagent grade, ≥99.0% (titration), crystalline
Sigma-Aldrich
Trizma® hydrochloride, BioPerformance Certified, suitable for cell culture, ≥99.0% (titration)
Sigma-Aldrich
Sodium chloride, Molecular Biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Sodium dodecyl sulfate, ReagentPlus®, ≥98.5% (GC)
Sigma-Aldrich
Sodium chloride, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
L-Leucine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
Sodium dodecyl sulfate, BioReagent, suitable for electrophoresis, Molecular Biology, ≥98.5% (GC)