DATA: Cancer-Associated Exosomes: Interactions with Glyconanotherapeutics and Their Impact on Exosome Integrity
Authors/Creators
- Wrobel, Dominika (Researcher)1
- Strašák, Tomáš (Researcher)2
- Bojarová, Pavla (Researcher)3
- Kocholatá, Michaela (Researcher)1
- Vrbata, David (Researcher)3
- Müllerová, Monika (Researcher)2
- Kurfiřt, Martin (Researcher)2
- Červenková Šťastná, Lucie (Researcher)2
- Mikšátko, Jiří (Researcher)4
- Malý, Jan (Researcher)1
- et al. Show all 11 authors
- Wrobel, Dominika (Researcher)1
- Strašák, Tomáš (Researcher)2
- Bojarová, Pavla (Researcher)3
- Kocholatá, Michaela (Researcher)1
- Vrbata, David (Researcher)3
- Müllerová, Monika (Researcher)2
- Kurfiřt, Martin (Researcher)2
- Červenková Šťastná, Lucie (Researcher)2
- Mikšátko, Jiří (Researcher)4
- Malý, Jan (Researcher)1
- Janoušková, Olga (Researcher)1
- 1. Centre of Nanomaterials and Biotechnology, Faculty of Natural Science Jan Evangelista University in Ústí and Labem, Pasteurova 15, Ústí and Labem, CZ-40096 Czech Republic
- 2. Institute of Chemical Process Fundamentals of the Czech Academy of Sciences, Rozvojova 1/135, CZ-16500 Praha 6, Czech Republic
- 3. Laboratory of Biotransformation, Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, CZ-14200, Praha 4, Czech Republic
- 4. Imaging Methods Core Facility, Charles University, Faculty of Science, BIOCEV, Vestec 252 50, Czech Republic
Description
Dataset contains data related to Interactions of Cancer-Associated Exosomes with Glyconanotherapeutics and Their Impact on Exosome Integrity.
Methods (English)
Exs were isolated from cells cultured in 175 cm3 flasks in 25 ml of medium. After 24 h of cultivation, the medium was replaced by Ex-free DMEM/DMEM-F12 (Gibco). Cells were cultivated for 48 h, then Exs were isolated using an ultracentrifuge (Beckman Coulter's OPTIMA XPN 90). Briefly, the medium was passed through a 0.22 μm filter and centrifuged at 2000 x g for 10 min at 4 oC to pellet the cell debris, then the supernatant was carefully transferred to a new tube and centrifuged at 10 000 x g for 20 min at 4 oC. The supernatant was then ultracentrifuge at 100 000 x g for 70 min at 4 oC. The pellets of Exs were washed with PBS and repeatedly ultracentrifuged at 100 000 x g for 70 min at 4 oC. Then the pellet of Exs was suspended in 300-500 µL of phosphate buffer saline (PBS) supplemented with cOmplete protease inhibitor cocktail (8 mM, Roche, Merck) or 300 µL RIPA lysis for western blot analysis. The protein concentration was measured using a BCA Protein Assay Kit (Pierce, ThermoFisher) and number of Exs were evaluated using Nanoparticle Tracking Analysis (NanoSight NS 3000, Malvern Instruments,UK). Samples were stored at -20 °C until further analysis.
Notes (English)
The research was supported from the project NU23-08-00307 by the Ministry of Health of the Czech Republic, from the projects UJEP-SGS-2022-53-005-3 and CZ.02.01.01/00/22_008/0004562. P.B. acknowledges support from the project 22-00317S by the Czech Science Foundation and from the mobility project LUC23148 by the Ministry of Education, Youth and Sports of the Czech Republic. D. V. acknowledges support from the Czech Academy of Sciences, Prospective Human Resources Support Program 2023 (project L200202352). The authors acknowledge Imaging Methods Core Facility at BIOCEV, institution supported by the MEYS CR (LM2023050 Czech-BioImaging) for their support & assistance in this work. The authors acknowledge Imaging Methods Core Facility at BIOCEV, institution supported by the MEYS CR (LM2023050 Czech-BioImaging) for their support & assistance in this work. The authors thank Jakub Červený from the Institute of Microbiology, Czech Academy of Sciences, for the determination of affinities by ELISA assays.