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Analiza krvne slike in proliferacija krvnih celic v kulturi pri proteusu
Prša, Patrik (Author), Bizjak Mali, Lilijana (Mentor) More about this mentor... This link opens in a new window, Sessions, Stanley K. (Co-mentor)

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Abstract
The purpose of this study was to develop non-destructive approaches to study cell biology, immunology and physiology in the European blind cave salamander (Proteus anguinus). Because of certain unique biological characteristics and its subterranean habitat in permeable limestone, the proteus is thought to be extremely vulnerable to different pollutants and to various potential pathogens that are of global concern to amphibians. Thus, developing non-destructive in vitro systems for research on the proteus is potentially an important approach in the conservation biology of this charizmatic species. Cell cultures in particular could provide an important source of material for a wide range of studies and can be done without sacrificing the animal or disrupting populations. We used small samples of blood to perform hematological analyses, including differential blood cell counts and neutrophil/lymphocyte ratios (NLR). We found that blood cell counts and differential WBC (white blood cell) counts did not change significantly in captivity and no statistically significant differences were found in the NLR between recently captured animals and animals kept in captivity for different lengths of time. Performing the same hematological analyses in diseased specimens likewise revealed no consistent pattern. Comparison of same individuals during different lengths of captivity showed no significant changes. We also developed two non-destructive cell culture methods: blood cell cultures and primary tissue explants from tail fin. Although our attempts to optimize blood cell cultures were only partially successful, we were able to optimize the protocol for primary tissue explants. Our results allow us to describe the dynamics of in vitro cellular migration and cellular lifespan for the first time in the proteus.

Language:Slovenian
Keywords:blood, hematology, cell culture, tissue explants, European blind cave salamander, Proteus anguinus
Work type:Master's thesis/paper (mb22)
Organization:BF - Biotechnical Faculty
Year:2018
COBISS.SI-ID:4879439 Link is opened in a new window
Views:439
Downloads:344
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Secondary language

Language:English
Title:Analysis of the blood count and blood cell proliferation in culture of proteus
Abstract:
Namen raziskave je bil razviti nedestruktven pristop za raziskave celične biologije, imunologije in fiziologije pri jamski repati dvoživki proteusu ali človeški ribici (Proteus anguinus). Zaradi nekaterih edinstvenih bioloških značilnosti in podzemnega habitata v propustnem apnencu je proteus izredno občutljiv in izpostavljen potencialnim onesnažilom in različnim patogenom, ki so globalna ogrožaja za dvoživke. Zaradi tega je razvijanje nedestruktivnih in vitro sistemov pomemben pristop za raziskave na proteus in ohranitev te karizmatične živalske vrste. Celične kulture so lahko pomemben vir materiala za številne študije in jih je mogoče izvesti brez žrtvovanja živali ali motenja populacij. Na majhnih vzorcih krvi smo izvedli hematološke analize, vključno z diferencialno belo krvno sliko (WBC) in razmerjem nevtrofilov in limfocitov (NLR), ki je pokazatelj fiziološkega stresa živali. Ugotovili smo, da se krvna slika in diferencialna bela krvna slika v ujetništvu bistveno ne spreminjata, prav tako ni bilo statistično značilnih razlik v NLR med živalmi iz narave in živalmi, ki so bile različno dolgo zadrževane v ujetništvu. Hematološka analiza pri bolnih osebkih prav tako ni razkrila nikakršnega značilnega vzorca. Primerjava istih osebkov po različnih časih v ujetništvu tudi ni pokazala razlik. Razvili smo tudi dve nedestruktivni metodi celične kultivacije: krvne celice in eksplantate repnega tkiva. Čeprav so bili naši poskusi optimizacije krvnih celic le delno uspešni, smo uspešno optimizirali postopek tkivnih eksplantov in opisali dinamiko celične migracije in življenjsko dobo celic in vitro pri proteusu.

Keywords:kri, hematologija, celične kulture, tkivni eksplanti, repata jamska dvoživka, Proteus anguinus

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