uPA and MMP‐2 were involved in self‐assembled network formation in a two dimensional co‐culture model of bone marrow stromal cells and endothelial cells

Abstract

Two dimensional (2D) co-cultures of human bone marrow stromal cells (HBMSCs) and human umbilical vein endothelial cells (HUVECs) stimulate osteoblastic differentiation of HBMSCs, induce the formation of self-assembled network and cell interactions between the two cell types involving many vascular molecules. Because of their strong activities on angiogenesis and tissue remodeling, urokinase plasminogen activator (uPA), plasminogen activator inhibitor-1 (PAI-1), matrix metalloproteinase-2 (MMP-2) as well tissue inhibitors of matrix metalloproteinase-2 (TIMP-2) were investigated in this 2D co-culture model. We found that the expression of uPA, MMP-2 in the co-cultured cells was significantly higher than those in mono-cultured cells. In opposite, PAI-1, expressed only by HUVECs is not regulated in the co-culture. Inhibition assays confirm that uPA played a critical role in the formation of self-assembled network as neutralization of uPA disturbed this network. In the same context, inhibition of MMP-2 prevented the formation of self-assembled network, while the inhibition of uPA abolished the over expression and the activity of MMP-2. This upregulation could initiate the uPA expression and proteolysis processes through the MMP-2 activity, and may contribute to endothelial cell migration and the formation of this self-assembled network observed in these 2D co-cultured cells.

1a,25-Dihydroxycholecalciferol (Vitamin D3) Induces NO-Dependent Endothelial Cell Proliferation and Migration in a Three-Dimensional Matrix

“…D and NO stimulation. MMP-2 involvement in PAE cells migration well correlated with previous studies reporting a constitutive MMP-2 expression in endothelium [27,29], and its upregulation during endothelial cells migration and tube formation in a 3D matrix [30]. …”

“…In particular, the main components of vascular basal lamina (collagen IV, laminin and fibronectin), are degraded mainly by MMP-2 and MMP-9, also known as gelatinases [29]. In particular, MMP-2 has received growing attention as it is the main MMP involved in angiogenesis [30]. …”

Cell interactions between human progenitor-derived endothelial cells and human mesenchymal stem cells in a three-dimensional macroporous polysaccharide-based scaffold promote osteogenesis

“…HBMSCs and HUVEC in a 2-D co-culture have been shown to up-regulate MMP-2 expression in the self-assembled network formed when HUVEC and HBMSCs were co-cultured together on plastic culture dishes, although the active form MMP-2a was undetectable, even though the MMP-2 inhibitor I prevented the selfassembled network observed in 2-D co-culture [70]. In these 3-D cultures, MMP-2 was found in its active form.…”

Improved vasculogenesis and bone matrix formation through coculture of endothelial cells and stem cells in tissue‐specific methacryloyl gelatin‐based hydrogels

“…The positive effect of osteogenic media supplements on vasculogenic EC behavior was observed in coculture with MSCs on cell culture plastic (Pedersen et al, ). Increased expression of VEGF by osteoblasts (Grellier et al, ) and increased expression of proteolytic enzymes important for EC migration (Li, Daculsi, Bareille, Bourget, & Amedee, ; Li et al, ) were also seen when culturing ECs and MSCs together in two‐dimensional (2D) culture. In addition, the 3D cell culture is more and more used because the 3D environment results in a more native cell behavior compared with 2D culture (Tibbitt & Anseth, ).…”