No public access
master's thesis
Bv-2 cells as an alternative model for the study of cytomegalovirus infekction of microglia

Marina Marcelić (2016)
University of Rijeka
Department of Biotechnology
Metadata
TitleBv-2 stanice kao alternativni model za izučavanje citomegalovirusne infekcije mikroglije
AuthorMarina Marcelić
Mentor(s)Natalia Kučić (thesis advisor)
Abstract
BV-2 stanice predstavljaju imortaliziranu mikroglijalnu staničnu liniju kao najčešće korištenu zamjenu za primarnu kulturu mikroglija stanica. Uobičajeno je da se uzgajaju u DMEM (engl. Dulbecco’s Modified Eagle Medium) mediju uz dodatak fetalnog goveĎeg seruma (FCS, engl. Fetal Calf Serum). In-vitro stanični mikrookoliš ne moţe dati točnu presliku onoga u središnjem ţivčanom sustavu, što naposljetku moţe dovesti do morfoloških i funkcionalnih promjena u mikroglija stanicama. Mikroglija stanice po prirodi su reaktivne i stoga su podloţne promjenama u svojoj okolini. Početni eksperimenti koje smo proveli u našem laboratoriju na spomenutoj liniji, u svrhu uspostavljanja in-vitro modela infekcije, pokazali su slabu permisivnost BV-2 mikroglija stanica za mišji citomegalovirus (MCMV). TakoĎer BV-2 mikroglija stanice pokazivale su obiljeţja stalne aktiviranosti i to u M1 aktivacijskome stanju. Kako bi utvrdili mogu li promjene u uvjetima kultivacije utjecati na permisivnost ovih stanica za MCMV, kao i na njihova fenotipska obiljeţja, stanice su uzgajane u različitim postocima FCS-a (0%, 2% i 5%) te su inficirane MCMV-om. Rezultati ovoga rada pokazuju da manipulacija uvjetima kultivacije, posebice oduzimanje FCS-a iz medija za uzgoj, dovodi do morfoloških promjena BV-2 mikroglija stanica. TakoĎer je povećana permisivnost bipolarnih BV-2 mikroglija stanica za MCMV infekciju. Različiti uvjeti uzgoja utjecali su i na razinu izraţaja proupalnih biljega iNOS, CD86 i CD16/32 te protuupalnog biljega CD206, ovisno o koncentraciji FCS-a. Zaključujemo da okolišne promjene koje mijenjaju metabolički i imunološki profil mogu dovesti do promijenjenog odgovora BV-2 mikroglija stanica na patogene.
Keywordsmicroglial BV-2 cell line FCS murine cytomegalovirus susceptibility to MCMV infection M1/M2 activation state
Parallel title (English)Bv-2 cells as an alternative model for the study of cytomegalovirus infekction of microglia
Committee MembersMiranda Mladinić Pejatović (committee chairperson)
Igor Jurak (committee member)
Natalia Kučić (committee member)
GranterUniversity of Rijeka
Lower level organizational unitsDepartment of Biotechnology
PlaceRijeka
StateCroatia
Scientific field, discipline, subdisciplineBIOTECHNICAL SCIENCES
Biotechnology
Study programme typeuniversity
Study levelgraduate
Study programmeDrug research and development
Academic title abbreviationmag. pharm. inv.
Genremaster's thesis
Language Croatian
Defense date2016-07-20
Parallel abstract (English)
Murine microglial BV-2 cell line is commonly used as a substitute for primary microglia. They are usually cultivated in the Dulbecco’s modified eagle medium (DMEM) supplemented with fetal calf serum (FCS). This in vitro cellular environment is unable to imitate the one present in the central nervous system, thus causing morphological and functional changes to the microglial cells. Microglial cells are reactive by nature, and as such are affected by all changes in their environment. In the first experiments we performed with the goal of establishing an in-vitro infection model, BV-2 microglial cells exhibited low susceptibility to murine cytomegalovirus (MCMV). Also, they exhibited features of a permanent M1 activity. To determine whether the changes in the conditions of cultivation may affect the susceptibility of these cells to MCMV infection as well as their phenotypic characteristics, the cells were cultivated in varying FCS percentage (0%, 2% and 5%), while also being infected with MCMV. Results from our study show that manipulation of cultivation conditions, specifically FCS depletion, leads to morphological changes of BV-2 microglial cells. Also, results show increased susceptibility of rod-shaped BV-2 microglial cells for MCMV infection. Varying FCS depletion also affected the levels of pro-inflammatory markers iNOS, CD86 and CD16/32 and the antinflammatory marker CD206, depending of the concentration. This indicates that the environmental changes that modulate the metabolic and immunological profile can cause an altered response of microglial cells to pathogens.
Parallel keywords (Croatian)BV-2 mikroglijalna stanična linija FCS mišji citomegalovirus permisivnost za MCMV infekciju M1/M2 aktivacijsko stanje
Resource typetext
Access conditionNo public access
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:193:372272
CommitterIvana Dorotić