20.500.12556/DKUM-52851 Kulture človeških endotelnih celic, izoliranih iz arterije mamarije interne Human endothelial cell cultures isolated from internal mammary artery Izhodišča: Cilj raziskave je pripraviti homogene kulture endotelnih celic (EC), izoliranih iz človeške arterije mamarije interne (AMI), dobljene med srčno obvodno operacijo, in jih vzdrževati v dolgoročnih kulturah. Na ta način bi skušali ugotovili pogoje, pri katerih bi lahko v nadaljnjem delu s celičnimi kulturami prišli do zadostnega števila avtolognih arterijskih EC celic za uporabo pri populaciji, ki jo kardiovaskularne bolezni najbolj prizadenejo. Metode: V raziskavo je bilo vključenih 7 moških, starih 52-76 let, z diagnosticirano ishemično srčno boleznijo. Pri vseh je bila opravljena obvodna operacija. Iz žilnih vzorcev, ki smo jih pridobili med operacijo, smo osamili celice na dva načina: s proteolitično razgradnjo tkiva in situ ali pa s proteolitično razgradnjo iz predhodno razkosanih žilnih preparatov. Spremljali smo morfologijo in kinetiko rasti EC v 3 tedne starih kulturah. Celično specifičnost smo dokazali z imunohistokemičnim določanjem von Willebrandovega faktorja. Rezultati: Rast celic je bila prisotna le v kulturah, ki smo jih pripravili s prvo metodo. Celice so kazale tipične morfološke značilnosti EC in izražale von Willebrandov faktor. Največje število celic v kolonijah (v povprečju 11, maksimalno pa 51) so kulture dosegle v 6 dneh. Podvojitven čas celic je bil 82+-43 ur. Zaključki: Z raziskavo smo pokazali, da je predstavljena metoda osamitve EC iz majhnega vzorca AMI primerna za pridobivanje vitalnih celic. Metoda je selektivna EC in omogoča njihovo rast v dolgoročnih kulturah. Končno število celic v kolonijah je za zdaj premajhno za uporabo v tkivnem inženirstvu, vendar bi se z optimizacijo metode v prihodnjih raziskavah lahko bolj približali temu cilju. Background: The aim of our study was to prepare homogenic and long tern cultures of endothelial cells (EC) from a human internal mammary artery (IMA) obtained during a cardiac by pass graft operation (CABG). In this way we would be able to assess the conditions suitable for the preparation of sufficient number of autologous arterial ECs and their use in a population mostly affected with cardiovascular diseases. Methods: Seven 52-76 years old male patients diagnosed with ischemic heart disease were included in ourstudy. Samples of internal mammary artery were collected during CABG performed on these patients. EC cells were prepared from these samples using two approaches. in situ digestion with proteolytical enzymes and proteolytic digestion from the previously resected IMA vessels. Cell morphology and growthkinetics were studied in 3 weeks old cultures. EC cell specificy was determined by immunohystochemical staining for von Willebrandćs factor. Results: Cell growth could be observed only in cultures prepared with the in situ proteolytic digestion. Cells sowed the typical EC morphology and were positive for von Willebrand's factor. The maximal cell number in colonies (11 in average, 51 at the most) was reached after 6 days in culture. The average cell doubling time of the five cultures was 82+-43 hours. Conclusions: The study proved that our method of isolation is specific for ECs. Using a small IMA sample we were able to collect a number of colonies and sustain them in long term cultures, but the end numbers of cells is still too small for their application in tissue engineering (TE). Our further efforts will be focused onthe optimization of the conditions which would allow sufficient proliferation of EC so that small IMA sample would be enough for the TE. endotelne celice celične kulture arterija mamarija interna tkivno inženirstvo endothelial cell cell cultures internal mammary artery tissue ingeneering true false true Slovenski jezik Angleški jezik Znanstveno delo 2015-07-10 18:32:12 2015-07-10 18:32:12 2022-06-01 03:07:43 2021-05-10 03:29:35 2006 0 0 str. 621-628 št. 10 Letn. 75 2006 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1318-0347 61 32893696 8913180 1318-0347 URN:SI:UM:DK:SOOSHWTG Zdravniski_vestnik_2006_Jagric_et_al._Kulture_cloveskih_endotelnih_celic,_izoliranih_iz_arterije_mamarije_interne.pdf Zdravniski_vestnik_2006_Jagric_et_al._Kulture_cloveskih_endotelnih_celic,_izoliranih_iz_arterije_mamarije_interne.pdf 1 00DFD0762FF1A57BC5EB51C95B7C4B5E fc4a17b97182468c9964e3d0dec2b64c750b7b983d94e9df2c6090f9741215bb 06dcac9e-7c0e-11eb-bb7a-00155d0001ca https://dk.um.si/Dokument.php?lang=slv&id=110236 http://vestnik.szd.si/index.php/ZdravVest/article/view/2033 1 https://dk.um.si/Dokument.php?lang=slv&id=110237 Ekonomsko-poslovna fakulteta 0 0 0