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2 protocols using β cyclodextran

1

Photodynamic Therapy for Breast Cancer

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Dopamine hydrochloride, ammonia aqueous solution (NH4OH, 28–30% wt%), Chlorin e6 (Ce6) and cell uptake inhibitors (nocodazole, cytochalasin D, chlorpromazine, genistein, and β-cyclodextran) were purchased from Sigma-Aldrich (Shanghai, China). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was provided by KeyGEN Biotech. Co., Ltd. (Jiangsu, China). Amine-PEG-thiol (NH2-PEG-SH, MW ≈ 2000 Da) was obtained from Laysan Bio. Inc. (Shanghai, China). Singlet oxygen sensor green (SOSG) kits were purchased from Invitrogen (Shanghai, China). Human breast tumor cells (SKBR-3) and human mammary epithelial cells (MCF 10A) were purchased from ATCC (Manassas, VA). Fetal bovine serum (FBS), trypsin, media and phosphate buffered saline (PBS) were purchased from Gibco/Life Technologies (Shanghai, China).
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2

Culturing and Maintaining Helicobacter pylori

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Strains and plasmids used in this study are listed in Table 1. Unless noted otherwise, H. pylori strains were grown as previously described (Carpenter et al., 2015 (link)). Briefly, H. pylori stock cultures were maintained at −80°C in H. pylori freezing media [brain heart infusion broth (BD Biosciences) containing 10% fetal bovine serum (FBS) and 20% glycerol (EMD chemicals, Inc)]. Freezer stocks were plated on horse blood agar (HBA) plates comprised of 4% Columbia agar (Neogean Corporation), 5% defibrinated horse blood (HemoStat Laboratoris, Dixon, CA), 2 mg/mL β-cyclodextran (Sigma), and an antibiotic/antifungal cocktail [10 μg/ml vancomycin (Amresco), 5 μg/ml cefsulodin (Sigma), 2.5 U/ml polymyxin B (Sigma), 5 μg/ml trimethoprim (Sigma), and 8 μg/ml amphotericin B (Amresco)]. Following growth on HBA plates, H. pylori strains were grown in liquid culture. Liquid media consisted of brucella broth (Neogen Corporation) supplemented with 10% FBS (Gibco) and 10 ug/mL vancomycin. Where indicated, 25 μg/mL Kanamycin (Kan25) was used to supplement the media. All cultures were grown at 37°C, in gas evacuation jars, under microaerobic conditions (5% O2, 10% CO2, and 85% N2) generated with an Anoxomat gas evacuation and replacement system (Advanced Instruments, Inc.); in addition, liquid cultures were grown shaking at 100 rpm.
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