20.500.12556/RUL-185832 Izolacija membranskih mikroveziklov Isolation of membrane microvesicles Membranski mikrovezikli sodelujejo pri prenosu biološko aktivnih molekul in imajo s tem pomembno vlogo pri medcelični komunikaciji ter uravnavanju imunskih in regenerativnih procesov. Njihova sestava odraža izvor in fiziološko oziroma patološko stanje celic, zato predstavljajo obetavne biomarkerje za zgodnje odkrivanje bolezni, spremljanje napredovanja raka in ocenjevanje uspešnosti zdravljenja. Preko njihove sestave lahko razberemo tudi ciljno celico, h kateri so namenjeni. Pri različnih boleznih se spreminjata tako koncentracija kot sestava membranskih mikroveziklov. Posamezni proteinski in drugi označevalci na veziklih, zato omogočajo razlikovanje med različnimi bolezenskimi stanji in napovedovanje njihovega izida. Kljub velikemu diagnostičnemu in terapevtskemu potencialu ostaja njihova izolacija pomemben izziv zaradi heterogenosti veziklov, prisotnosti netarčnih delcev in pomanjkanja standardiziranih metod. Med najpogosteje uporabljenimi postopki izolacije so ultracentrifugiranje, ultrafiltracija in izključitvena kromatografija. Zadnja leta pa se uveljavljajo tudi druge metode, ki ponujajo ugodnejšo alternativo. Mednje sodijo tangencialna filtracija v kombinaciji z izključitveno kromatografijo, ultracentrifugiranje s pretočno citometrijo, šaržna ultrafiltracija in mikrofluidika, ki jo je potrebno še nadgraditi za uporabo na večjem nivoju in ločevanje subpopulacij zunajceličnih veziklov. Pri tem kombinacija metod v večini raziskav zagotavlja boljšo čistost, izkoristek, ločitev subpopulacij in kakovost izoliranih mikroveziklov. Potrebna bo predvsem optimizacija karakterizacije, da bodo rezultati izolacije mikroveziklov zanesljivejši. Nadaljnje raziskave bodo ključne za standardizacijo izolacijskih postopkov ter širšo uporabo membranskih mikroveziklov v diagnostiki in zdravljenju. Membrane microvesicles mediate the transfer of biologically active molecules, thus play a crucial role in intercellular communication as well as in the regulation of immune and regenerative processes. Their molecular composition reflects both the cellular origin and the physiological or pathological state of the parent cells, making them potential biomarkers for the early detection of disease, monitoring cancer progression, and evaluating treatment succes. Furthermore, their molecular composition can provide insight into the target cells to which they are directed. In various diseases, both the concentration and molecular composition of membrane microvesicles, including specific protein and other molecular markers, are altered, enabling detection between different pathological conditions and prediction of disease outcomes. Despite their considerable diagnostic and therapeutic potential, the isolation of membrane microvesicles remains a major challenge due to vesicle heterogeneity, the presence of non-target particles, and the lack of standardized isolation protocols. The most commonly used isolation methods include ultracentrifugation, ultrafiltration, and size-exclusion chromatography. In recent years, additional techniques have been developed to improve the specificity of isolated products. Among the most promising techniques are tangential flow filtration combined with size-exclusion chromatography, ultracentrifugation with flow cytometry, batch ultrafiltration, and microfluidic technologies which still requires further optimization to enable large-scale application and efficient separation of extracellular vesicle subpopulations. In most studies, combining multiple isolation methods has resulted in improved purity, higher yield, sepparation of subpopulations and quality of isolated microvesicles. Further optimisation of characterization methods is also required to improve reliability of isolation results. Continued research will be essential for the standardization of isolation protocols and the broader implementation of membrane microvesicles in diagnostic and therapeutic applications. Membranski mikrovezikli zunajcelični vezikli izolacija ektosomi Membrane microvesicles extracellular vesicles isolation ectosomes true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2026-08-21 08:15:33 2026-08-21 08:15:46 2026-08-22 04:19:29 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 285316 288452099 60332.pdf 60332.pdf 1 8E4F5833342729C8B425133C70DEF228 fafe0601dbf74f7488cabbf960ab065b81c1b4df5ea6cb0b1ec7098cde528981 5b5d726f-9d27-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=244821 Biotehniška fakulteta 0 0 0