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3 protocols using 154 cf culture medium

1

Murine Basal Cell Carcinoma In Vitro

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RB, fetal bovine serum (FBS), penicillin/streptomycin solution, trypsin-EDTA solution, ABDA probe [9,10-antherachenediyl-bis(methylene) dimalonic acid], MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide], and HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) were purchased from Sigma-Aldrich (Taufkirchen, Germany). Dulbecco’s phosphate buffered saline without calcium and magnesium (DPBS) was purchased from Biowest (Nuaillé, France). HBSS (Hanks’ Balanced Salt Solution) and 154 CF culture medium were obtained from Gibco/ThermoFisher Scientific (Waltham, MA, USA). Chelex 100 Resin was obtained from Bio-Rad (Hercules, CA, USA). H2DCFDA (2′,7′ dichlorodihydrofluorescein diacetate) was purchased from Invitrogen/ThermoFisher Scientific (Waltham, MA, USA). Dimethyl sulfoxide (DMSO) was purchased from POCH (Gliwice, Poland). Murine basal cell carcinoma lines (AsZ, BsZ, CsZ) were provided by Dr. Ervin Epstein (Children’s Oakland Research Institute, Oakland, CA, USA).
Organic solvents were dried and freshly distilled under argon prior to use. Chemicals were obtained from commercial sources and used as received. Water solutions of chemicals, amphiphiles and buffer solutions were prepared using milliQ® deionized water. Sonication of amphiphile solutions was done in a Sonorex Super RK 31 H ultrasonic bath (Bandelin Electronic, Berlin, Germany).
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2

Cellular Oxidative Stress Evaluation

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The RB, fetal bovine serum
(FBS), penicillin/streptomycin solution, trypsin–EDTA solution,
ABDA probe [9,10-antherachenediyl-bis(methylene) dimalonic acid],
MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide],
and HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) were
purchased from Sigma-Aldrich (Taufkirchen, Germany). Dulbecco’s
phosphate-buffered saline without calcium and magnesium (DPBS) was
purchased from Biowest (Nuaillé, France). HBSS (Hanks’
balanced salt solution) and the 154 CF culture medium were obtained
from Gibco/ThermoFisher Scientific (Waltham, MA, USA). Chelex 100
Resin was obtained from Bio-Rad (Hercules, CA, USA). H2DCFDA (2′,7′-dichlorodihydrofluorescein diacetate)
was purchased from Invitrogen/ThermoFisher Scientific. Dimethyl sulfoxide
(DMSO) was purchased from POCH (Gliwice, Poland). Murine basal cell
carcinoma lines (AsZ, BsZ, and CsZ) were provided by Dr. Ervin Epstein
(Children’s Oakland Research Institute, Oakland, CA, USA).
Poly(propyleneimine) (PPI) dendrimers of the third and fourth generationa with 32 or 64 primary amino surface groups, respectively,
were obtained from Symo-Chem (Eindhoven, the Netherlands). Poly(amidoamine)
(PAMAM) dendrimers of the third and fourth generation with 32 and
64 primary amino surface groups, respectively, were obtained from
Sigma-Aldrich.
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3

Synthesis of Phosphorus Dendrimers and Conjugates

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Phosphorus dendrimers (and their conjugates with RB) of the first, second and third generation were synthesized in the Laboratoire de Chimie de Coordination, CNRS (Figure 11).
Rose bengal, tyramine, cesium carbonate, thionyl chloride, 1-(2-aminoethyl)pyrrolidine, triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), fetal bovine serum, penicillin/streptomycin solution, trypsin–EDTA solution, and MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) were purchased from Sigma-Aldrich, Taufkirchen, Germany and TCI, Zwijndrecht, Belgium. Dulbecco’s phosphate buffered saline with no calcium and no magnesium (DPBS) was purchased from Biowest, France. 154 CF culture medium was obtained from Gibco, Thermo Fisher Scientific, Waltham, MA, USA. Chelex 100 Resin was obtained from Bio-Rad. Singlet oxygen sensor green (SOSG) was purchased from Molecular Probes, ThermoFisher, Waltham, MA, USA. Dimethyl sulfoxide (DMSO) was purchased from POCH (Gliwice, Poland). Snake-Skin™ Dialysis Tubing, 3.5K MWCO, 22 mm was obtained from ThermoFisher, Waltham, MA, USA. Murine basal cell carcinoma lines (AsZ, BsZ, CsZ) were kindly provided by Dr. Ervin Epstein (Children’s Oakland Research Institute, Oakland, CA, USA).
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