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2 protocols using flasks and multiwell plates for in vitro studies

1

Doxorubicin-Dendrimer Nanocarrier for Breast Cancer

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All chemical reagents were purchased from commercial suppliers. Solvents for the synthesis were purchased from Sigma-Aldrich. All cell culture reagents were purchased from Gibco® (Germany). Flasks and multiwell plates for in vitro studies were obtained from Nunc (Germany). Amine terminated PAMAM G4 dendrimer, doxorubicin hydrochloride, PBS (phosphate buffered saline), FBS (fetal bovine serum) and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) were purchased from Sigma-Aldrich. Trypan blue was purchased from Molecular Probes (USA). Caelyx® (PEGylated liposomal doxorubicin) was purchased from Janssen-Cilag International (USA). Rubusoside and kaempferol were purchased from Sigma-Aldrich. Human breast adenocarcinoma cell line MCF-7 (ATCC no. HTB-22) was purchased from ATCC (USA). AntiGLUT1-FITC antibody (FAB1418F) was purchased from R&D.
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2

PAMAM-Doxorubicin Conjugate Evaluation

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All chemical reagents were purchased from commercial suppliers. Solvents for the synthesis were purchased from Sigma-Aldrich (Poznan, Poland). All cell culture reagents were purchased from Gibco® (Life Technologies Polska Sp. z o. o., Warsaw, Poland). Flasks and multiwell plates for in vitro studies were obtained from Nunc (Life Technologies Polska Sp. z o. o., Warsaw, Poland). Amine terminated PAMAM G4 dendrimer, doxorubicin hydrochloride, PBS (phosphate buffered saline), FBS (fetal bovine serum) and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) were purchased from Sigma-Aldrich. Trypan blue was purchased from Molecular Probes (Thermo Scientific™, Warsaw, Poland). Herceptin (trastuzumab) was a gift from Roche Poland. Human breast adenocarcinoma’s cell lines: HER-2 negative (MCF-7 ATCC no. HTB-22) and HER-2 positive (SKBR-3 ATCC no. HTB-30) were purchased from ATCC (LGC Standards Sp. z o. o., Lomianki, Poland).
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