Mesenchymal Stem Cells Support Migration, Extracellular Matrix Invasion, Proliferation, and Survival of Endothelial Cells In Vitro

We investigated effects of the paracrine factors secreted by human mesenchymal stem cells (hMSCs) on endothelial cell migration, extracellular matrix invasion, proliferation, and survival in vitro. Human mesenchymal stem cells were cultured as a monolayer or as three‐dimensional aggregates in hanging drops (hMSC spheroids). We performed analysis of paracrine factors in medium conditioned by a monolayer of hMSCs and hMSC spheroids. Concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor, angiogenin, procathepsin B, interleukin (IL)‐11, and bone morphogenic protein 2 were increased 5–20 times in medium conditioned by hMSC spheroids, whereas concentrations of IL‐6, IL‐8, and monocyte hemoattractant protein‐1 were not increased. Concentrations of VEGF and angiogenin in medium conditioned by hMSC spheroids showed a weak dependence on the presence of serum, which allows serum‐free conditioned medium with elevated concentrations of angiogenic cytokines to be obtained. Medium conditioned by hMSC spheroids was more effective in stimulation of umbilical vein endothelial cell proliferation, migration, and basement membrane invasion than medium conditioned by a monolayer of hMSCs. This medium also promotes endothelial cell survival in vitro. We suggest that culturing of hMSCs as three‐dimensional cellular aggregates provides a method to concentrate proangiogenic factors secreted by hMSCs and allows for reduction of serum concentration in conditioned medium. Our data support the hypothesis that hMSCs serve as trophic mediators for endothelial cells.

[1]  S. Gerson,et al.  Phenotypic and functional comparison of cultures of marrow‐derived mesenchymal stem cells (MSCs) and stromal cells , 1998, Journal of cellular physiology.

[2]  Hans Hauner,et al.  Cartilage-like gene expression in differentiated human stem cell spheroids: a comparison of bone marrow-derived and adipose tissue-derived stromal cells. , 2003, Arthritis and rheumatism.

[3]  H. Ohgushi,et al.  Intravenous administration of mesenchymal stem cells improves cardiac function in rats with acute myocardial infarction through angiogenesis and myogenesis. , 2004, American journal of physiology. Heart and circulatory physiology.

[4]  M. Burnett,et al.  Marrow-Derived Stromal Cells Express Genes Encoding a Broad Spectrum of Arteriogenic Cytokines and Promote In Vitro and In Vivo Arteriogenesis Through Paracrine Mechanisms , 2004, Circulation research.

[5]  A. Caplan,et al.  Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5‐azacytidine , 1995, Muscle & nerve.

[6]  X. Qin,et al.  Paracrine action enhances the effects of autologous mesenchymal stem cell transplantation on vascular regeneration in rat model of myocardial infarction. , 2005, The Annals of thoracic surgery.

[7]  V. Goldberg,et al.  The Chondrogenic Potential of Human Bone-Marrow-Derived Mesenchymal Progenitor Cells* , 1998, The Journal of bone and joint surgery. American volume.

[8]  Martin Fussenegger,et al.  Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. , 2003, Biotechnology and bioengineering.

[9]  Yi Li,et al.  Gliosis and brain remodeling after treatment of stroke in rats with marrow stromal cells , 2005, Glia.

[10]  M G Nichols,et al.  Oxygen diffusion and reaction kinetics in the photodynamic therapy of multicell tumour spheroids. , 1994, Physics in medicine and biology.

[11]  A I Caplan,et al.  Characterization of cells with osteogenic potential from human marrow. , 1992, Bone.

[12]  R. Kloner,et al.  Allogeneic Mesenchymal Stem Cell Transplantation in Postinfarcted Rat Myocardium: Short- and Long-Term Effects , 2005, Circulation.

[13]  M. Long,et al.  Three-dimensional cellular development is essential for ex vivo formation of human bone , 2000, Nature Biotechnology.

[14]  U. Losert,et al.  Bone marrow stromal cells can provide a local environment that favors migration and formation of tubular structures of endothelial cells. , 2005, Tissue engineering.

[15]  J. Ingwall,et al.  Evidence supporting paracrine hypothesis for Akt‐modified mesenchymal stem cell‐mediated cardiac protection and functional improvement , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[16]  M. Chopp,et al.  Intravenous bone marrow stromal cell therapy reduces apoptosis and promotes endogenous cell proliferation after stroke in female rat , 2003, Journal of neuroscience research.

[17]  A. Kosaki,et al.  Implantation of Bone Marrow Mononuclear Cells Into Ischemic Myocardium Enhances Collateral Perfusion and Regional Function via Side Supply of Angioblasts, Angiogenic Ligands, and Cytokines , 2001, Circulation.

[18]  David M. Bodine,et al.  Bone marrow cells regenerate infarcted myocardium , 2001, Nature.

[19]  M. Burnett,et al.  Local Delivery of Marrow-Derived Stromal Cells Augments Collateral Perfusion Through Paracrine Mechanisms , 2004, Circulation.