Vpliv encimskega ekstrakta riževe semenske ovojnice na rast, razvoj in preživetje mezenhimskih matičnih celic, izoliranih iz maščobnega tkiva podgane

Škof, Maja (author), Marc, Janja (mentor), Cano Rodriquez, Maria Mercedes (co-mentor)

Encimski ekstrakt riževe ovojnice (EERB) je produkt encimske hidrolize riževe ovojnice z endoproteazami. Je vodotopna zmes peptidov, polisaharidov in polinenasičenih maščobnih kislin, ki je bogata z antioksidanti (γ-orizanol, vitamin E). Mezenhimske matične celice, izoliranih iz maščobnega tkiva podgane (ADSC) so multipotentne matične celice, ki predstavljajo potencialen vir celic v regenerativni medicini. Njihova izolacija je relativno enostavna, prav tako imajo širok potencial za diferenciacijo, zato jih mnoge študije uporabljajo kot vir za regeneracijo različnih tkiv. V magistrski nalogi smo izvedli preliminarne študije, da bi ugotovili, kako EERB učinkuje na različne lastnosti ADSC. Zanimalo nas je, ali EERB kot dodatek mediju za gojenje celic vpliva na proliferacijo ADSC, diferenciacijo ADSC v nevrosfere in nevrone, ter ali EERB kot antioksidant pomaga zaščiti ADSC pred vplivom oksidativnega stresa. Pokazali smo, da je dodatek EERB celičnemu mediju učinkovito sredstvo za boljšo rast celic. S primerno koncentracijo v celičnem mediju EERB spodbuja rast ADSC, kar smo ugotovili s testom MTS in štetjem celic v Neubauerjevi celici. Pokazali smo tudi, da EERB vpliva na izražanje genov ADSC na primeru SOX2 in eEF. Ena izmed potencialnih uporab ADSC v regenerativni medicini je diferenciacija teh celic v nevrone. Proces diferenciacije je težaven in močno odvisen od okolja, kjer se celice nahajajo. Zaradi svojih antioksidativnih lastnosti bi EERB lahko vplival na bolj učinkovito diferenciacijo ADSC, čeprav je za dokončne zaključke potrebno še nekaj raziskav. Prav tako nas je zanimalo, ali EERB kot antioksidant varuje celice pred oksidativnim stresom. Z MTS testom in štetjem celic v Neubauerjevi komori smo pokazali, da je proliferacija celic v okolju z oksidativnim stresom bolj uspešna, če so celice pred tem izpostavljene EERB. Zanimivo je, da je LDH test citotoksičnosti pokazal, da je CH v kombinaciji s predhodno izpostavitvijo celic z EERB bolj toksičen kot če ekstrakta ne uporabimo. Tudi v tem primeru bi bilo potrebno narediti dodatne raziskave. Zaključimo lahko, da EERB ima potencial za nadaljnje raziskave in bi lahko vplival na bolj uspešno proliferacijo in diferenciacijo ADSC tudi v okoljih, kjer je prisoten oksidativni stres.

• Language: Slovenian
• Keywords: Ključne besede: mezenhimske matične celice, encimski ekstrakt riževe ovojnice, proliferacija, oksidativni stres.
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2024
• PID: 20.500.12556/RUL-154795
• Publication date in RUL: 02.03.2024

Secondary language

• Language: English
• Title: Effect of enzymatic extract from rice bran on growth, differentiation and survival of rat adipose tissue-derived mesenchymal stem cells

Abstract

Enzymatic extract of rice bran (EERB) is water-soluble extract that exerts strong antioxidant activity. It is obtained after enzyme hydrolysis of rice bran with endoproteases. It contains a mixture of water-soluble substances, rich in antioxidants (γ-oryzanol, vitamin E) and consists mainly of peptides, polysaccharides and polyunsaturated fatty acids. Adipose tissue-derived mesenchymal stem cells (ADSC) are multipotent stem cells, which are a prominent source in regenerative medicine. Because of their relatively easy isolation and broad differentiation potential, many studies are focusing on different tissue regeneration with ADSC. However, many difficulties in their growth and differentiation still exist. This Master’s thesis is a preliminary study where we evaluated the effect of EERB on different characteristics of ADSC. We researched how EERB as a cell culture medium influences proliferation of ADSC, their differentiation to neurospheres and neurons and if EERB as an antioxidant protects ADSC against oxidative stress, induces by cumene hydroperoxide (CH). EERB as a cell culture medium supplement has shown to be beneficial for growth of ADSC. In adequate concentrations EERB promotes cell growth, as we have shown with MTS assay and cell counting experiments. Also, EERB does influence ADSC gene expression, as we researched for SOX2 and eEF. In regenerative medicine, ADSC are of great interest for differentiation to neurons, though the process is arduous and strongly depended on environment and cell conditions. EERB could be an important factor in more efficient differentiation because of its antioxidative features though more research should be done on this aspect. Additionally, we showed that EERB protects ADSC against oxidative stress, that was induced by CH. However, more research on this topic is needed. The results of MTS assay and cell counting showed that EERB does protect against oxidative stress, but LDH cytotoxicity assay was showing stronger cytotoxicity when cells were incubated with CH and EERB. We can conclude that EERB is indeed a promising new factor for cells proliferation and its influence should be considered in future research.

• Keywords: Key words: adipose tissue-derived mesenchymal stem cells, enzymatic extract from rice bran, cell proliferation, oxidative stress.