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2 protocols using foetal bovine serum fbs

1

Isolation and Culture of Human Skin Fibroblasts

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All recruited individuals signed an informed consent form approved by our Institutional Ethics Committees prior to being enrolled in the study on a voluntary basis (Ethical research board of the CHU de Québec. Protocol number: 2012–1316. For more information, please contact (gurecherche@chuq.qc.ca). All experiments were performed in compliance with the national Tri-Council Policy Guidelines: Ethical Conduct for Research Involving Humans and approved by the ethics committee of the CHU de Quebec – Université Laval. Fibroblasts were isolated from skin biopsies of 6 informed consent healthy individuals (Supplementary Table 5) as previously described33 (link). Fibroblasts were grown in Dulbecco-Vogt modification of Eagle’s medium (DMEM; Invitrogen, Burlington, ON, Canada) supplemented with 10% foetal bovine serum (FBS; VWR, Radnor, PA, USA), 100 IU/mL penicillin G (Sigma-Aldrich, Oakville, QC, Canada), and 25 μg/mL gentamicin (Schering, Pointe-Claire, QC, Canada). Cell cultures were kept in an incubator at 37 °C, 8% CO2 and 95% relative humidity, and media was changed every 2 days.
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2

Bioactive Compound Acquisition and Preparation

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Kaempferol (K) and luteolin (L) were purchased from Indofine (Hillsborough, New Jersey, USA). Tamarixetin (T), (-)-epicatechin (EC), (-)-gallocatechin (GC), (-)-epicatechin gallate (ECG) and (-)-epigallocatechin gallate (EGCG) were purchased from Extrasynthese (Genay, France). Quercetin (Q), isorhamnetin (I), Dimethyl sulfoxide (DMSO), Mc Coy's 5A medium, poly-DL-alanine and green tea catechins (GTC) mix (containing (+)-catechin (C), (-)-catechin gallate (CG), EC, ECG, EGCG, GC and (-)-gallocatechin gallate (GCG), 100 µg/mL of each component delivered in acetonitrile: water (8:2) with 5% 1 M HCl) were purchased from Sigma-Aldrich (Schnelldorf, Germany). Penicillin, streptomycin, Dulbecco's phosphate buffered saline (DPBS) and GlutaMax were purchased from Gibco TM (Life Technologies, Paisley, United Kingdom). Foetal bovine serum (FBS) was purchased from Lonza TM (VWR, Verviers, Belgium). Ascorbic acid was purchased from Prolabo TM (VWR, Leuven, Belgium). Acetonitrile (ACN) and formic acid (UPLC-MS grade) were purchased from BioSolve BV (Valkenswaard, The Netherlands). High-purity water was produced with a Synergy UV water purification system from Merck Millipore (Amsterdam, The Netherlands).
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