Jing Lin1, Merry L Lindsey, Beili Zhu, C Mauli Agrawal, Steven R Bailey. 1. Janey Briscoe Center for Cardiovascular Research, Division of Cardiology, Department of Medicine, University of Texas Health Science Center at San Antonio, TX 78229-3900, USA. linj2@uthscsa.edu
Abstract
PURPOSE: Adipose-derived stromal cells (ADSCs) have been shown to increase angiogenesis in ischemic tissue. Maintaining cell survival and facilitating angiogenesis in ischemic tissue, however, continues to be the major challenge of ADSCs implantation. Recently, bioengineered scaffolds were introduced to support and facilitate cell culture and differentiation. The effects of a surface modified three-dimensional (3D) scaffold on ADSC function have not been investigated. Accordingly, the objective of this study was to determine the influence of a gas-plasma treated scaffold on ADSC growth, differentiation into endothelial cell, and angiogenic gene expression. METHODS: Freshly isolated mouse ADSCs were characterized by flow cytometry and cultured into wells containing gas-plasma treated scaffolds, non-treated scaffolds, or control wells. Either endothelial growth media or differentiation media was used to alter cell environment. After 3 and 6 days, cell proliferation was analyzed. VEGF concentration in the medium was measured by ELISA. Gene expression was quantified by real-time PCR for VEGF receptor-2 (KDR), cyclooxygenase-2 (COX-2) and matrix metalloproteinases-2 (MMP-2). RESULTS: ADSCs expressed stem/endothelial progenitor markers CD34 and CD133 and endothelial cell marker CD31. ADSCs grew in the 3D scaffold. Cells grown on gas-plasma treated scaffolds displayed significantly increased expression of VEGF, COX-2, and MMP-2 when grown in differentiation but not growth media. When cultured in endothelial growth media, VEGF secretion and the expression of KDR, COX-2 and MMP-2 were lower in 3D scaffolds than controls. CONCLUSIONS: This study suggests that 3D scaffolds, especially gas-plasma treated scaffolds, support ADSC growth and support differentiation into endothelial cells.
PURPOSE:Adipose-derived stromal cells (ADSCs) have been shown to increase angiogenesis in ischemic tissue. Maintaining cell survival and facilitating angiogenesis in ischemic tissue, however, continues to be the major challenge of ADSCs implantation. Recently, bioengineered scaffolds were introduced to support and facilitate cell culture and differentiation. The effects of a surface modified three-dimensional (3D) scaffold on ADSC function have not been investigated. Accordingly, the objective of this study was to determine the influence of a gas-plasma treated scaffold on ADSC growth, differentiation into endothelial cell, and angiogenic gene expression. METHODS: Freshly isolated mouse ADSCs were characterized by flow cytometry and cultured into wells containing gas-plasma treated scaffolds, non-treated scaffolds, or control wells. Either endothelial growth media or differentiation media was used to alter cell environment. After 3 and 6 days, cell proliferation was analyzed. VEGF concentration in the medium was measured by ELISA. Gene expression was quantified by real-time PCR for VEGF receptor-2 (KDR), cyclooxygenase-2 (COX-2) and matrix metalloproteinases-2 (MMP-2). RESULTS: ADSCs expressed stem/endothelial progenitor markers CD34 and CD133 and endothelial cell marker CD31. ADSCs grew in the 3D scaffold. Cells grown on gas-plasma treated scaffolds displayed significantly increased expression of VEGF, COX-2, and MMP-2 when grown in differentiation but not growth media. When cultured in endothelial growth media, VEGF secretion and the expression of KDR, COX-2 and MMP-2 were lower in 3D scaffolds than controls. CONCLUSIONS: This study suggests that 3D scaffolds, especially gas-plasma treated scaffolds, support ADSC growth and support differentiation into endothelial cells.