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The in vitro effects of antibiotics on cell viability and gene expression of equine bone marrow-derived mesenchymal stromal cells

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Original languageEnglish
Pages (from-to)355-360
Number of pages6
JournalEquine Veterinary Journal
Volume44
Issue number3
DOIs
StatePublished - 1 May 2012

Abstract

Objectives: To investigate the effects of commonly used antibiotics on cell viability and gene expression of equine bone marrow-derived mesenchymal stromal cells (MSC) in vitro. Materials and methods: Bone marrow-derived MSC were cultured in media containing gentamicin, amikacin, penicillin, enrofloxacin or ceftiofur at concentrations of 50, 100, 200 and 500μg/ml. The alamarBlue fluorescence assay was used to assess cell viability over 48 h. After 5 days the cells were released and lysed prior to RNA extraction and reverse transcription. RNA levels were assessed using spectrophotometry and quantitative PCR was used to analyse gene expression of COL1A2, COL5A1, TNC, TNFα, CASP3, BCl2 and TGFβR2 relative to the reference gene GAPDH. Results: Enrofloxacin produced a significant concentration-dependent reduction in cell viability at 200μg/ml and higher concentrations (P = 0.009). Amikacin significantly reduced cell viability at 500μg/ml (P = 0.002). Penicillin had no effect on cell viability at the concentrations tested (P = 0.18). Gentamicin and ceftiofur showed some interaction with the assay but had no overall effect on cell viability. At 500μg/ml gentamicin (P

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