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Optimizacija izolacije zunajceličnih veziklov iz plazme in cerebrospinalne tekočine
ID Šilc, Tina (Author), ID Videtič Paska, Alja (Mentor) More about this mentor... This link opens in a new window

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Abstract
Zunajcelični vezikli so majhne membransko zaprte strukture, ki se sproščajo iz pravzaprav vseh celic. Vsebujejo različne vrste molekul, vključno z majhnimi RNA in so udeleženi v številne fiziološke procese, zaradi česar se vse bolj preiskuje njihova vloga potencialnih biooznačevalcev, ki bi zdravnikom lahko pripomogli pri prilagajanju strategij zdravljenja posameznih pacientov in jim pomagali prepoznati tiste, pri katerih je večja verjetnost, da se bodo odzvali na določen pristop k zdravljenju. Vendar pa standardizacija in nadaljnja karakterizacija tehnik ločevanja, ki se uporabljajo pri študijah zunajceličnih veziklov, še vedno še ni bila izvedena. S tem namenom smo testirali nekaj izmed sodobnih molekularno-bioloških metod za izolacijo zunajceličnih veziklov iz cerebrospinalne tekočine in plazme pri tem pa sledili smernicam MISEV. Kot primerna metoda za izolacijo zunajceličnih veziklov iz cerebrospinalne tekočine se je izkazalo ultracentrifugiranje, za izolacijo zunajceličnih veziklov iz plazme pa tekočinska kromatografija za hitro ločevanje beljakovin v povezavi z ultracentrifugiranjem ter ultracentrifugiranje na saharozni blazinici, vendar ne gre za najbolj optimalni metodi, zato bo potrebna še nadaljnja optimizacija.

Language:Slovenian
Keywords:zunajcelični vezikli, plazma, cerebrospinalna tekočina, izolacija, samomord
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2022
PID:20.500.12556/RUL-134650 This link opens in a new window
ISSN:
COBISS.SI-ID:96233219 This link opens in a new window
Publication date in RUL:24.01.2022
Views:1574
Downloads:193
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Secondary language

Language:English
Title:Optimisation of extracellular vesicles isolation from plasma and cerebrospinal fluid
Abstract:
Extracellular vesicles are small membrane-enclosed structures that are released from practically all cells. They contain different types of molecules, including small RNAs, and are involved in many physiological processes, which has led to increasing research into their role as potential biomarkers that could help doctors tailor treatment strategies to individual patients and help them identify those who are more likely to respond to a particular treatment approach. However, standardisation and further characterisation of the separation techniques used in extracellular vesicle studies is still ongoing. To this end, we have tested some of the current molecular biological methods for the isolation of extracellular vesicles from cerebrospinal fluid and plasma, following the MISEV guidelines. Fast protein liquid chromatography Ultracentrifugation was found to be a suitable method for isolation of extracellular vesicles from cerebrospinal fluid, and fast protein liquid chromatography in conjunction with ultracentrifugation and sucrose gradient centrifugation were considered to be suitable methods for isolation of extracellular vesicles from plasma, but given the results, further optimisation will be needed.

Keywords:extracellular vesicles, plasma, cerebrospinal fluid, isolation, suicide

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