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Permeabilnost izbranih učinkovin skozi spontano transformirano Caco-2 celično linijo s sposobnostjo hitre vzpostavitve integritete = Permeability of selected substances through a self-transforming Caco-2 cell line with rapid integrity development : enoviti magistrski študijski program FarmacijaKristan, JercaMonosloj celične linije Caco-2 se pogosto uporablja kot in vitro model za določanje humane instestinalne absorpcije. Referenčni protokol predvideva 21±1 dni, da celice po nasajanju na filtre dosežejo ... ustrezno stopnjo diferenciacije in ustvarijo konfluenten monosloj, ki se nato lahko uporabi za permeabilnostni poskus. Slabost Caco-2 celic je zato ravno izredno dolg čas kultivacije. V celičnem laboratoriju Fakultete za farmacijo se je z naraščajočim številom pasaž pojavil sev celične linije Caco-2, ki je začel izkazovati lastnosti pospešene diferenciacije in sposobnost pospešene vzpostavitve integritete, zaradi česar doseže ustrezno konfluenten monosloj veliko prej. V naši nalogi smo ta sev ovrednotili po genotipizaciji s strani ATCC, ki je sev prepoznala kot avtentično Caco-2 celično linijo; izoblikovali smo protokol za gojenje, s katerim smo želeli ugotoviti, kako gostota nasaditve celic na inserte vpliva na hitrost deljenja in čas do vzpostavitve integritete. To smo preverjali z merjenjem TEER ter s preverjanjem permeabilnosti modelnih spojin. Po tem protokolu smo 5. dan od nasaditve celic na filtre za gojenje dobili pripravljen monosloj za meritve permeabilnosti in primerjavo z referenčnimi, nespremenjenimi Caco-2 celicami. Na podlagi smernic FDA smo izoblikovali tri skupine modelnih spojin, ki so omogočale sočasno kvantifikacijo v akceptorskih raztopinah, z njimi pa smo ovrednotili ustreznost ločevanja med dobro in srednje ter slabo permeabilnimi spojinami. Primerjali smo pridobljene rezultate med referenčnim in testiranim sevom, ki je izkazoval sposobnost hitre vzpostavitve integritete ter potrdili, da testirani sev hitreje doseže konfluenten monosloj, uporaben za določanje permeabilnosti; ugotovili smo, da ima testiran sev na podlagi merjenja TEER in prepustnosti FITC-dekstranov ustrezno celično integriteto, da dobro ločuje dobro permeabilnime spojine od srednje in slabo permeabilnih ter da izraža dobro korelacijo med humano absorpcijo in vivo ter permeabilnostjo učinkovin in vitro.Type of material - master's thesis ; adult, seriousPublication and manufacture - Ljubljana : [J. Kristan], 2022Language - slovenianCOBISS.SI-ID - 128961283
Author
Kristan, Jerca
Other authors
Žakelj, Simon
Topics
celice Caco-2 |
permeabilnost |
absorpcija učinkovin |
integriteta |
Lucifer Yellow |
analitske metode
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Library/institution |
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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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Kristan, Jerca | ![]() |
Žakelj, Simon | 22659 |
Source: Personal bibliographies
and: SICRIS
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