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Vpeljava postopka za določanje koncentracije mezenhimskih matičnih celic v bioloških vzorcih
Klemen, Tjaša (Author), Maličev, Elvira (Mentor) More about this mentor... This link opens in a new window

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Abstract
V zadnjem desetletju postajajo mezenhimske matične celice (MMC) zaradi sposobnosti diferenciacije v različne celice in obsežnega parakrinega delovanja vse bolj uporabne v regenerativni medicini. Uspešno so bile izolirane iz skoraj vseh vrst tkiv, vendar problem predstavlja njihova maloštevilčnost, in pomankanje standardiziranih postopkov za določanje koncentracije MMC v sveže izoliranih vzorcih. Namen naše raziskave je bil vpeljati postopek za štetje MMC v vzorcih kostnega mozga, stimulirane periferne krvi in maščobnega tkiva s pretočnim citometrom, z uporabo protiteles proti celičnim označevalcem CD271, CD45 in fluorescentnih kroglic za natančno štetje celic. Po vpeljavi postopka smo naše rezultate uspešno primerjali z rezultati testa določanja števila fibroblastov, ki tvorijo kolonije (CFU-F). V vzorcih smo vzporedno določali tudi koncentracijo krvotvornih matičnih celic. Največjo koncentracijo MMC smo določili v vzorcih maščobnega tkiva, najmanjšo v vzorcih periferne krvi. Za razliko od koncentracije krvotvornih matičnih celic in števila CFU-F je bila korelacija med koncentracijo MMC in starostjo darovalcev vzorcev statistično neznačilna, vendar bi bilo potrebno v nadaljno raziskavo vključiti večje število in starostno bolj raznolike darovalce.

Language:Slovenian
Keywords:matične celice, mezenhimske matične celice, kostni mozeg, periferna kri, maščobno tkivo
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2018
Publisher:[T. Klemen]
UDC:602.9:611.018(043.2)
COBISS.SI-ID:8973433 Link is opened in a new window
Views:847
Downloads:179
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Secondary language

Language:English
Title:The implementation of the procedure for determination of mesenchymal stem cells concentration in biological samples
Abstract:
In the last decade Mesenchymal Stem Cells (MSC) are being used more extensively in regenerative medicine. This is due to their ability to differentiate into various cells and due to their extensive paracrine function. They have been successfully isolated from almost every type of tissue, the problem, however, lies in their small numbers and the lack of standardised procedures to determine MSC concentration in freshly isolated samples. The aim of our research was to introduce a procedure for counting MSC in the samples of bone marrow, stimulated peripheral blood and fat tissue with a flow cytometer, using antibodies against cell markers CD271, CD45 and florescent beads for precise cell count. After introducing the procedure, we successfully compared our results with the test determining the count of colony-forming units - fibroblasts (CFU-F). At the same time, we used the samples to determine the concentration of haematopoietic progenitor cells. The highest concentration of MSC was found in the sample of fat tissue, the lowest in those of peripheral blood. Unlike the concentration of haematopoietic progenitor cells and the count of CFU-F, the corelation between the concentration of MSC and the age of the donor samples was statistically insignificant. To get more conclusive results, we should expand our research to a larger group of donors from many different age groups.

Keywords:stem cells, mescenchymal stem cells, bone marrow, peripheral blood, fat tissue

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