Oxidative stress is associated with a wide variety of pathologies. An adequate model of oxidative damage of cells is important for the research of the effects of substances on such damaged cells.
Fibroblast is a basic cell of connective tissue, which plays a critical role in wound healing. 3T3 cells are a standard fibroblast cell line obtained from Swiss albino mouse embryo tissue.
The aim of this study was to design and development of in vitro model of oxidative damage of 3T3 cell line. As a relevant pathophysiological source of ROS components of the Weisberger biogenic oxidative system [Cu(II), ascorbate] were selected (Baňasová et al., 2014). Indication of ROS presence was performed by using flurescent dyes (dichlorofluorescin diacetate and dihydroethidium).
This work was supported by grant VEGA 2/0065/15.
M. Baňasová, K. Valachová, J. Rychlý, I. Janigová, K. Csomorová, R. Mendichi, D. Mislovičová, I. Juránek, L. Šoltés (2014) Effect of Bucillamine on Free-Radical-Mediated Degradation of High-Molar-Mass Hyaluronan Induced in vitro by Ascorbic Acid and Cu(II) Ions. Polymers 6, 2625-2644; doi:10.3390/polym6102625.
otazka
Dobry den, velmi ma zaujala vasa praca, preto by som sa rada spytala jednu otazku.
Akym sposobom budete hodnotit vplyv, predpokladam, testovanych latok na tento in vitro model? Chystate sa kvantifikovat obrazky z fluorescencneho mikroskopu, alebo robit spektrofluorimetricke merania? Vopred dakujem za odpoved
Re: otazka
Re: otazka
Dobry den, dakujem velmi pekne za zaujem o moju pracu.
Co sa tyka prvej otazky, vplyv testovanych latok (dalej len latka) budem hodnotit dvoma sposobmi:
1. Latka sa prida pred vyvolanim oxidacneho stresu, aby som mohla pozorovat, ci dokaze bunky pred poskodenim ochranit.
2. Latka sa prida po vyvolani oxidacneho stresu (cca 15 minut), ale este pred vymenou media, tymto sposobom chcem sledovat vplyv latok na uz poskodene bunky a ci ich dana latka dokaze "zachranit". V tomto smere uz su aj nejake vysledky (ktore vynikli az po zaciatku konferencie).
Co sa tyka kvantifikacie ROS (obrazky z fluorescencneho mikroskopu), mam v plane tuto kvantifikaciu pomocou cytometru.