Karakterizacija matičnih ćelija izolovanih iz zubne pulpe mlečnih zuba dece

  • Jasmina Debeljak Martačić Institute for Medical Research, University of Belgrade, Belgrade, Serbia
  • Jelena Francuski Department of Pathophysiology, Faculty of Veterinary Medicine, University of Belgrade, Belgrade, Serbia
  • Tijana Lužajić Department of Pathophysiology, Faculty of Veterinary Medicine, University of Belgrade, Belgrade, Serbia
  • Nemanja Vuković Faculty of Stomatology, University of Business Academy, Pančevo, Serbia
  • Slavko Mojsilović Institute for Medical Research, University of Belgrade, Belgrade, Serbia
  • Neda Drndarević Faculty of Stomatology, University of Business Academy, Pančevo, Serbia
  • Marijana Petakov Faculty of Stomatology, University of Business Academy, Pančevo, Serbia
  • Marija Glibetić Institute for Medical Research, University of Belgrade, Belgrade, Serbia
  • Danica Marković Department of Pathophysiology, Faculty of Veterinary Medicine, University of Belgrade, Belgrade, Serbia
  • Anita Radovanović Department of Pathophysiology, Faculty of Veterinary Medicine, University of Belgrade, Belgrade, Serbia
  • Vera Todorović Faculty of Stomatology, University of Business Academy, Pančevo, Serbia
  • Milica Kovačević Filipović Department of Pathophysiology, Faculty of Veterinary Medicine, University of Belgrade, Belgrade, Serbia
DOI: https://doi.org/10.2298/3715
Ključne reči: dental pulp||, ||zub, pulpa, stem cells||, ||ćelije, matične, tooth, deciduous||, ||denticija, mlečna, child, preschool||, ||deca, predškolska, cell differentation||, ||ćelija, diferencijacija, adipogenesis||, ||adipogeneza, chondrogenesis||, ||hondrogeneza, osteogenesis||, ||osteogeneza,

Sažetak


Uvod/Cilj. Prošla dekada je bila posebno obeležena naporima na polju korišćenja ex vivo razvijenih i usmeravanih mezenhimskih matičnih ćelija (MSCs), kao sredstva za različite tipove regenerativne terapije. Cilj ove studije bio je da se utvrdi imunofenotip i potencijal za proliferaciju i diferencijaciju ćelija izolovanih iz zubne pulpe mlečnih zuba dece eksfoliranih u periodu kada je koren zuba bio u poslednjoj fazi resorpcije. Metode. Primarna adherentna populacija ćelija poreklom od pet donora dobijena je metodom eksplanta. Prisustvo progenitorskih ćelija koje obrazuju kolonije fibroblasta (CFU-F) pokazano je u prvoj pasaži. Do treće pasaže ćelije su ekspandirane, a potom korišćene za analiziranje. Imunofenotip je određen korišćenjem protočne citometrije. Proliferativni potencijal i vreme udvajanja ćelija (PDT) u kulturi je definisano na osnovu apsolutnog broja ćelija na početku i na kraju svake pasaže. Posle tronedeljne kultivacije ćelija u komercijalnim medijumima za stimulaciju osteogeneze, hondrogeneze i adipogeneze, citološkim i histološkim metodama je određena morfologija ćelija i karakteristike vanćelijskog matriksa. Rezultati. Antigeni koji karakterišu mezenhimske matične ćelije CD29 i CD73 su bili eksprimirani na svim nediferenciranim ćelijama, dok su antigeni CD146 i CD106 bili eksprimirani na ograničenom broju ćelija. Antigen CD34 (karakterističan za ćelije hematopoetske loze) nije bio eksprimiran. Incidencija CFU-F bila je 4,7 ± 0,5/100 ćelija. PDT se nije menjao tokom osam pasaža i u proseku je iznosio 25 h. Posle tronedeljne stimulacije diferencijacije u kulturama sa adipogenim medijumom došlo je do stvaranja ćelija sa masnim kapljicama, a u kulturama sa osteogenim medijumom došlo je do formiranja vanćelijskog matriksa sa deponovanim kalcijumovim solima. U kulturama sa hondrogenim medijumom došlo je do stvaranja tkiva sličnog hrskavici i vanćelijskog matriksa sa glikozaminoglikanima i kolagenom II. Zaključak. Zubna pulpa mlečnih zuba dece sadrži ćelijsku populaciju koja odgovara mezenhimskim matičnim ćelijama prema svojim karakteristikama, ima visok proliferativni potencijal i potencijal da se diferencira u tri ćelijske linije što je čini pogodnom za dalje in vitro analize i evaluaciju ćelijske terapije.

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Objavljeno
2015/04/23
Broj časopisa
Vol. 71 Br. 8 (2014)
Rubrika
Originalni članak