20.500.12556/RUL-170407 Izražanje mlečnih proteinov v primarnih epitelnih celičnih kulturah goveje mlečne žleze Milk Protein Expression in Primary bovine mammary epithelial cells Mlečna žleza je organ, značilen za sesalce, katerega funkcija je tvorba in izločanje mleka, ki je vir hranil za potomce. Mleko poleg drugih komponent vsebuje tudi mlečne proteine, ki jih delimo na kazeine in proteine sirotke. Sinteza mlečnih proteinov poteka v alveolih mlečne žleze, natančneje v luminalnih celicah, ki so obdane z mioepitelnimi celicami in omogočajo iztis mlečnih komponent iz alveolov v mlečne vode. Uporabili smo primarne celične linije epitelnih celic mlečne žleze in jih stimulirali z lakotogenimi hormoni. Z mikroskopiranjem in imunofluorescenco smo opazovali spremembe v morfologiji celične kulture in izražanju označevalcev. Ugotovili smo, da laktogeni hormoni sprožijo celično diferenciacijo in vplivajo na morfologijo, ki ustreza morfologiji mlečne žleze v laktaciji. V celični kulturi smo potrdili prisotnost predvsem mioepitelnega in luminalnega epitelnega podtipa. Z metodo qPCR smo opazovali vpliv laktogenih hormonov na izražanje izbranih genov za mlečne proteine. Ugotovili smo, da se geni za mlečne proteine CSN1S1, CSN3 in LALBA izražajo tudi v pogojih in vitro in da stimulacija z laktogenimi hormoni lahko poveča njihovo izražanje. Povečanje izražanja izbranih genov smo preverjali tudi na proteinski ravni z metodo prenosa western, katera nam zaradi nizkih koncentracij mlečnih proteinov v celičnih lizatih in gojišču ni uspelo nedvoumno pokazati povečanja izražanja. Ugotovitve našega eksperimenta so lahko v pomoč pri nadaljnjih raziskavah biologije laktacije in vpliva laktogenih hormonov na celice mlečne žleze ter imajo potencialno uporabnost v živinoreji za povečanje prireje mleka in za sintezo mlečnih proteinov in vitro. The mammary gland is an organ specific to mammals that has the task of producing and secreting milk – a vital source of nutrients for the offspring. In addition to other components, milk also consists of milk proteins, which can be divided into caseins and whey proteins. Milk protein synthesis takes place in the alveoli of the mammary gland, more precisely in the luminal cells, which are surrounded by myoepithelial cells that enable the secretion of milk components from the alveoli into the milk ducts. Our aim was to establish primary mammary epithelial cell lines from bovine milk and investigate how stimulation with lactogenic hormones affects morphology and milk protein expression. Using microscopy and immunofluorescence, we observed changes in the morphology of the cell culture and the expression of markers. Our results suggest that lactogenic hormones induce cell differentiation and influence morphology in a manner consistent with the morphology of lactating mammary glands. We confirmed the presence of myoepithelial and luminal cell subtypes. Using qPCR, we analysed the effect of lactogenic hormones on the expression of milk protein genes. Some milk protein genes are also expressed in vitro, and lactogenic stimulation can increase their expression. This increase in gene expression was also assayed at the protein level by Western bloting – however, due to the low protein concentrations in cell lysates and the growth media, we could not definitively prove the increase in protein synthesis. The results of our experiment may be useful for further research in the field of lactation biology and the effect of lactogenic hormones on mammary gland cells. Our results could also be used in livestock breeding to increase milk production and in vitro milk protein synthesis. mlečni proteini laktogeni hormoni izražanje genov goveja mlečna žleza milk proteins lactogenic hormones gene expression bovine mammary gland true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2025-07-04 11:05:00 2025-07-04 11:05:07 2025-07-24 10:58:07 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 25631 243689731 Ferjancic_Lara_-_Izrazanje_mlecnih_proteinov_v_primarnih_epitelnih_celicnih_kulturah_goveje_mlec.pdf Ferjancic_Lara_-_Izrazanje_mlecnih_proteinov_v_primarnih_epitelnih_celicnih_kulturah_goveje_mlec.pdf 1 235A5D11B1D92E0B17B22D2E71944E3D 2d3fa165de8fbed9a10e9c176c16b5a40dd75aa73ceb34303e1d86f57d32208e 13dbba9b-58b4-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=212406 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0