Mesenchymal stem cells isolated from peripheral blood and umbilical cord Wharton's jelly

Abstract

Introduction. Mesenchymal stem cells (MSCs) are a promising tool for regenerative medicine, but due to the heterogeneity of their populations, different sources and isolation techniques, the characteristics defining MSCs are inconsistent. Objective. The aim of this study was to compare the characteristics of MSCs derived from two different human tissues: peripheral blood (PB-MSCs) and umbilical cord Wharton's Jelly (UC-MSCs). Methods. The PB-MSC and UC-MSC were isolated by adherence to plastic after gradient-density separation or an explant culture method, respectively, and compared regarding their morphology, clonogenic efficiency, proliferating rates, immunophenotype and differentiation potential. Results. MSCs derived from both sources exhibit similar morphology, proliferation capacity and multilineage (osteogenic, chondrogenic, adipogenic and myogenic) differentiation potential. Differences were observed in the clonogenic capacity and the immunophenotype, since UC-MSCs showed higher CFU-F (colony-forming units-fibroblastic) cloning efficiency, as well as higher embryonic markers (Nanog, Sox2, SSEA4) expression. When additional surface antigens were analyzed by flow cytometry (CD44, CD90, CD105, CD33, CD34, CD45, CD11b, CD235a) or immunofluorescent labeling (vimentin, STRO-1 and α-smooth muscle actin), most appeared to have similar epitope profiles irrespective of MSC source. Conclusion. The results obtained demonstrated that both MSCs represent good alternative sources of adult MSCs that could be used in cell therapy applications.

Uvod. Mezenhimske matične ćelije (MMĆ) su posebno značajne za regenerativnu medicinu. S obzirom na heterogenost njihovih populacija, raznovrsnost izvora i tehnika izolacije, ne postoje jedinstvena obeležja koja određuju MMĆ. Cilj rada. Cilj ove studije bila je uporedna analiza bioloških karakteristika MMĆ izolovanih iz periferne krvi (PK-MMĆ) i Vartonove (Wharton) sluzi pupčane vrpce (VSP-MMĆ). Metode rada. PK-MMĆ i VSP-MMĆ su izolovane na osnovu osobine da prianjaju na plastičnu podlogu, a upoređivane su njihove morfološke odlike, klonogeni i proliferativni kapacitet, imunofenotip i potencijal diferencijacije. Rezultati. Dobijeni rezultati pokazali su da oba tipa MMĆ imaju slične morfološke osobine, sličan proliferativni kapacitet i multipotentni potencijal diferencijacije u ćelije različitih mezenhimskih tkiva (koštanog, masnog, hrskavičavog i mišićnog). Razlike su primećene u klonogenom kapacitetu i imunofenotipu, budući da su VSP-MMĆ ispoljile veći kapacitet za formiranje CFU-F (engl. colony-forming unit-fibroblasts), kao i veću ekspresiju markera tipičnih za embrionalne matične ćelije (Nanog, Sox2, SSEA4). Ispitivanje površinskih antigena tipičnih za MMĆ (CD44, CD90, CD105, CD33, CD34, CD45, CD11b, CD235a) protočnom citometrijom i imunofluorescentno obeležavanje dodatnih mezenhimskih markera (Vimentin, STRO-1 i alfa-aktina glatkih mišićnih ćelija) nije ukazalo na razlike u ekspresiji ovih markera. Zaključak. Oba tipa MMĆ su pogodan alternativni izvor adultnih MMĆ koje bi se mogle koristiti u ćelijskoj terapiji.