RESUMO
Many controversial results exist when comparing mesenchymal stromal cells (MSCs) derived from different sources. Reasons include not only variables in tissue origin, but also methods of cell preparation or choice of expansion media which can strongly influence the expression and hence, function of the cells. In this short report we aimed to investigate the expression of the cell anchoring proteins desmoglein 2, desmocollin 3 and plakophilin 2 in early passage placenta-derived MSCs of fetal (fetal pMSCs) and maternal (maternal pMSCs) origins versus adult bone marrow-derived MSCs (bmMSCs) that were expanded and cultured under the same good manufacturing practice (GMP) conditions. Comprehensive gene expression microarray analysis profiling indicated differential expression of these genes in the different MSC-derived types with fetal pMSCs expressing the highest levels of PKP2, DSC3 and DSG2, followed by maternal pMSCs, while bmMSCs expressed the lowest levels. A higher expression of PKP2 and DSC3 genes in fetal pMSCs was confirmed by qRT-PCR suggesting neonatal increases in the expression of these desmosomal genes vs. adult MSCs. Intracellular desmocollin 3 and desmoglein 2 expression was observed by flow cytometry and cytoplasmic plakophilin 2 by immunofluorescence in all three MSC sources. These data suggest that fetal pMSCs, maternal pMSCs and bmMSCs may anchor intermediate filaments to the plasma membrane via desmocollin 3, desmoglein 2 and plakophilin 2.
Assuntos
Desmocolinas/genética , Desmogleína 2/genética , Perfilação da Expressão Gênica/métodos , Células-Tronco Mesenquimais/metabolismo , Placenta/metabolismo , Placofilinas/genética , Adulto , Células da Medula Óssea/metabolismo , Diferenciação Celular/genética , Proliferação de Células/genética , Células Cultivadas , Desmocolinas/metabolismo , Desmogleína 2/metabolismo , Feminino , Feto/citologia , Imunofluorescência , Células HeLa , Humanos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Placenta/citologia , Placofilinas/metabolismo , Gravidez , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Placenta-derived mesenchymal stromal cells (pMSCs) are a very attractive source of MSCs. In this short report we evaluated the expression of phenotypic markers from fetal and maternal pMSCs after exposure to myogenic medium commonly used to differentiate bone marrow MSCs (bmMSCs) to smooth muscle-like cells (SMCs). In order to reveal differences between these different MSC sources, cells were expanded and differentiated to elucidate whether this differentiation protocol facilitated efficient differentiation of SMCs from human pMSCs. We report that TGF-ß1, PDGF and ascorbic acid is not sufficient to produce SMCs from pMSCs.