The breast cancer cell lines MCF-7, BT474 and SKBR3 and the normal breast cell line MCF-10A were purchased from the American Type Culture Collection (Manassas, VA, USA). All tumor cells were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 3 mM L-glutamine, 50 µg/ml gentamicin (Biowhittaker Reagents; Lonza Group, Ltd., Basel, Switzerland) and 1% penicillin/streptomycin. MCF-10A cells were cultured in a mammary epithelial cell medium (cat. no. 7611, MEpiCM; ScienCell Research Laboratories, Inc., San Diego, CA, USA). MCF-7 is the typical human breast carcinoma cell, therefore, this cell line was chosen for further experiments. All cells were cultured at 37°C in 5% CO2.
Biowhittaker reagents
Manufactured by Lonza
Sourced in Switzerland, Belgium
Biowhittaker Reagents are a range of laboratory products designed for use in cell culture, molecular biology, and other life science applications. These reagents provide essential components and solutions required for various laboratory procedures. They are manufactured to meet the highest quality standards to support reliable and consistent results in research and development activities.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Mepicm, by ScienCell (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
Bt474, by American Type Culture Collection (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Penicillin streptomycin pen strep, by Thermo Fisher Scientific (1 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Biowest (1 mentions)
3 protocols using biowhittaker reagents
1
Breast Cancer Cell Line Cultivation
2
Fibroblast Response to Nutrient Famine
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The in vitro fibroblast experiment was described in more detail previously [9 (link)]. In short, fibroblasts were obtained by skin biopsies from five healthy participants of Dutch descent, of which one was male and four were female (mean age = 38.4 years, sd = 7.0) (see Table 1 ). All participants provided written informed consent. Fibroblasts were plated in two T25 flasks in Minimum Essential Medium (MEM) (Gibco®) with 15% fetal bovine serum (FBS)(Gibco®) and 1% penicillin-streptomycin PenStrep (Gibco®) and in an atmosphere of 95% atmospheric air and 5% CO2 at 37 °C (normal conditions). After reaching 70–80% confluence, the supernatant was removed and the cells were washed three times with phosphate buffered saline (PBS) (BioWhittaker® Reagents, Lonza). Next, one of the T25 flasks from each donor was cultured in the non-famine condition with Minimum Essential Medium (MEM) (Gibco®) supported with 15% FBS, while the other T25 flasks were cultured in only Minimum Essential Medium (MEM) as famine condition. After 72 h, cells were harvested from each flask and stored as cell pellet for DNA isolation.
3
Evaluating TiO2 Nanoparticle Effects on Rat Blood
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Blood specimens were collected from 6 intact WAG rats in K 2 EDTA Vacutainer test tubes (Guangzhou, China). K 2 EDTA was used to prevent blood clotting. The EU Directive 2010/63/EU on the protection of animals used for scienti c purposes was followed in this study.
At once, blood portions (50 µl) were incubated in 5 ml RPMI-1640 medium (Biowest, France) and 5% fetal bovine serum (BioWhittaker® reagents, Lonza, Belgium) for 24 h with 0-10-20-40-100-200 mg/L TiO 2 nanoparticles of both types. Conditions and parameters studied are shown in Fig. 1a.
The research was conducted in compliance with the EU Directive 2010/63/EU on the protection of animals used for scienti c purposes and Ukrainian legislation and was approved by the Commission on Ethics and Bioethics at Kharkiv National Medical University (Kharkiv, Ukraine, minutes #3 dated August 28, 2020).
At once, blood portions (50 µl) were incubated in 5 ml RPMI-1640 medium (Biowest, France) and 5% fetal bovine serum (BioWhittaker® reagents, Lonza, Belgium) for 24 h with 0-10-20-40-100-200 mg/L TiO 2 nanoparticles of both types. Conditions and parameters studied are shown in Fig. 1a.
The research was conducted in compliance with the EU Directive 2010/63/EU on the protection of animals used for scienti c purposes and Ukrainian legislation and was approved by the Commission on Ethics and Bioethics at Kharkiv National Medical University (Kharkiv, Ukraine, minutes #3 dated August 28, 2020).
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