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Kultiviranje mišjih proliferativnih enterocitov in spremljanje aktivnosti maltaze = Cultivating mouse proliferative enterocytes and measuring activity of intestinal maltase : diplomska nalogaZemljić, MajaVpeljali smo metodo kultiviranja misjih proliferativnih enterocitov in pripravili dolgozivo celièno kulturo misjih enterocitov. Zrtvovali smo sest misi (BALBžC) moskega spola, starih stiri mesece. ... Zrtvovali smo jih v stanju mirovanja, za odvzem tankega èrevesa. Odvzeli smo srednji del tankega èrevesa,predvidoma jejunum. Vsebino tankega èrevesa smo sprali s fiziolosko raztopino in s skarjami (skalpelom) iztisnili vsebino tankega èrevesa ter rumenkasto povrsinsko sluz. Preostali del tankega èrevesa smo v petrijevki razrezali na èim manjse kosme, jih razgradili s kolagenazo in jih kultivirali.Medij, v katerem smo kultivirali enterocite je bil kompletiran RPMI medij. Nato smo skozi pasaze (ena pasaza pomeni priblizno en teden kultiviranja) spremljali aktivnosti maltaze in saharaze. Dokazali smo, da sta maltaza in saharaza aktivni ze na stopnji proliferativnega enterocita. Tako smo dobili zanesljiv parameter za prouèevanje toksiènih vplivov na enterocite,ker so disaharidaze kot membranski encimi obèutljivi oznaèevalci zapatoloske spremembe na enterocitih.Koncentracijo proteinov v celiènem lizatuoziroma homogenatu smo doloèili po modificirani Lowry-jevi metodi s Folin - Ciocalteujevim fenolnim reagentom. Specifiène aktivnosti disaharidaz, maltaze in saharaze v homogenatu, smo doloèili po Dahlqvist-ovi metodi, kjer specifièno aktivnost maltaze doloèimo preko hitrosti hidrolize maltoze na dve molekuli glukoze in saharoze na molekulo glukoze in fruktoze. Kolièino sprosèene glukoze smo merili z glukoza - oksidazno metodo.Uspesno smo izpeljali metodo kultiviranja misjih proliferativnih enterocitov in dokazali, da sta maltaza in saharaza aktivni ze na stopnji proliferativnega enterocita.Razviti model kultiviranja enterocitov je primeren za prouèevanje citotoksiènih vplivov npr. gliadina pri celiakiji.Type of material - undergraduate thesisPublication and manufacture - Ljubljana : [Zemljić, M.], 2006Language - slovenianCOBISS.SI-ID - 1954161
Author
Zemljić, Maja
Other authors
Lukač-Bajalo, Jana
Topics
mišji enterociti |
celiakija |
encim maltaza |
tkivna kultura |
celična kultura
Library/institution |
City | Acronym | For loan | Other holdings |
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Faculty of Pharmacy, Ljubljana | Ljubljana | FFALJ |
reading room 1 cop.
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Zemljić, Maja | |
Lukač-Bajalo, Jana | 03573 |
Source: Personal bibliographies
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