Analysis of matrix metalloproteinase activity during differentiation of mesenchymal stem cells isolated from different tissues of one donor

Resumo

Mesenchymal stem cells established from bone marrow (FetMSC) and limb bud (M-FetMSC) of early human embryo, as well as spheroids derived these cells, were induced to undergo osteogenic and adipogenic differentiation. Differentiated cells exhibited the activity of metalloproteinase (MMP)-9, -2, and -1. Its activity was different in osteogenic and adipogenic cells, as well as in monolayer cultures (2D) and cell spheroids (3D). The direct correlation between the level of adipogenic differentiation and gelatinases MMP-9 and MMP-2 activities in both cell lines in 2D and 3D culture was shown. M-FetMSC cells in 2D culture 12 days in culture during showed low potential for adipogenesis and reduced activity of MMP-2 and MMP-9. The low level of adipogenic differentiation in 2D M-FetMSC culture was accompanied with increased MMP-1 activity and enhanced differentiation (3D culture) resulted in a significant increase of both MMP activities. MMP-1 activity varied oppositely. MMP-1 activity declined in 3D cultures with a higher level of adipogenic differentiation. The level of osteogenic differentiation was similar in both cell lines during 2D and 3D cultivation. MMP-1 and -9 activities in both cell lines were not associated with osteogenic differentiation. MMP-2 and MMP-2 activity in these cells remained unchanged. The results suggest MMP implication in FetMSC and М-FetMSC differentiation. The difference in MMP activities during the cell differentiation may be caused by variations in the microenvironment or ECM properties in 2D and 3D cultures.