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Kultivacija matiÄnih i progenitorskih Äelija hematopoeze iz kostne srži hrÄka
Hamster, a hibernating animal, is an important experimental model in research on the influence of hypothermia on different physiological processes. A simple procedure for cultivation and identification of hamster hematopoetic stem cells (HSC) and hematopoetic progenitor cells (HPC) is a premise for a successful investigation upon hypothermia effects on hematopoiesis. The aim of this work was to evaluate the utilization of commercially available methylcellulose media (MC) and recombinant mouse and human cytokines for hamster HSC and HPC assays, in order to enable further studies on these cells. Hamster bone marrow mononuclear cells (BMMNC) were plated in MC containing cytokines that support mouse or human HPC growth. Also, BMMNC were resuspended in cytokine supplemented liquid media and incubated for 5 weeks with a four day monitoring of viable cell number. We demonstrated that hamster hematopoietic progenitor cells committed for erythroid lineage and myeloid lineage successfully formed recognizable colonies in both mouse and human MC, while multipotent progenitor cells formed colonies only in mouse MC. We also defined conditions for the evaluation of hamster HSC activity in liquid cultures, based on continuous 5 weeks HSC proliferation. The obtained results verify the utilization of mouse specific MC for further research on hamster HPC biology during hypothermia.FizioloÅ”ka hibernacija u koju hrÄci ulaze prilikom izlaganja niskim temperaturama, Äini ove životinje zanimljivim eksperimentalnim modelom za ispitivanje hematopoeze u uslovima hipotermije. Preduslov za ovo ispitivanje je postojanje jednostavne metode za kultivaciju i identifikaciju hematopoetskih Äelija hrÄka. Cilj ovog rada je bio da se ispita moguÄnost kultivacije progenitorskih Äelija hematopoeze hrÄka u kompletnoj metil celulozi dizajniranoj za kultivaciju miÅ”ijih i humanih hematopoetskih Äelija, kao i da se odrede optimalni uslovi za kultivaciju matiÄnih Äelija hematopoeze hrÄka u teÄnoj kulturi. Mononuklearne Äelije kostne srži hrÄka su posaÄene u metil celulozu i u teÄnu kulturu. Oba medijuma su sadržala kombinacije rekombinantnih miÅ”ijih i/ili humanih citokina. Kolonije progenitorskih Äelija opredeljenih za mijelopoezu i opredeljenih za eritropoezu su se formirale u metil celulozi dizajniranoj za kultivaciju miÅ”ijih i humanih hematopoetskih Äelija, dok su se primitivnije kolonije sastavljene od oba tipa Äelija (mijeloidna i eritrocitna loza) formirale samo u metil celulozi dizajniranoj za kultivaciju miÅ”ijih hematopoetskih Äelija. Osim toga, populacija matiÄnih Äelija hematopoeze hrÄka je proliferisala u teÄnim kulturama tokom 5 nedelja bez znakova opadanja proliferativnog potencijala. Ova istraživanja pokazuju da se primenjene metode mogu uspeÅ”no koristiti za ispitivanje hematopoeze kod hrÄka