Proteomic Profiling of Tissue-Engineered Blood Vessel Walls Constructed by Adipose-Derived Stem Cells

Abstract

Adipose-derived stem cells (ASCs) can differentiate into smooth muscle cells and have been engineered into elastic small diameter blood vessel walls in vitro. However, the mechanisms involved in the development of three-dimensional (3D) vascular tissue remain poorly understood. The present study analyzed protein expression profiles of engineered blood vessel walls constructed by human ASCs using methods of two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS). These results were compared to normal arterial walls. A total of 1701 -15 and 1265 -26 protein spots from normal and engineered blood vessel wall extractions were detected by 2DE, respectively. A total of 20 spots with at least 2.0-fold changes in expression were identified, and 38 differently expressed proteins were identified by 2D electrophoresis and ion trap MS. These proteins were classified into seven functional categories: cellular organization, energy, signaling pathway, enzyme, anchored protein, cell apoptosis/defense, and others. These results demonstrated that 2DE, followed by ion trap MS, could be successfully utilized to characterize the proteome of vascular tissue, including tissue-engineered vessels. The method could also be employed to achieve a better understanding of differentiated smooth muscle protein expression in vitro. These results provide a basis for comparative studies of protein expression in vascular smooth muscles of different origin and could provide a better understanding of the mechanisms of action needed for constructing blood vessels that exhibit properties consistent with normal blood vessels.

RETRACTED ARTICLE: Small Diameter Blood Vessels Bioengineered From Human Adipose-derived Stem Cells

“…The mechanical properties of the bioengineered vessel were examined as previously described 13 . Briefly, the bioengineered vessels and normal human HSV with an inner diameter of 4 mm were sectioned circumferentially.…”

“…Several attempts have been made to utilize mature cells or different types of stem cells for blood vessel engineering. Approaches include human aortic smooth muscle cells (HASMCs) 12 , or human adipose-derived stem cells (hASCs) 13 . or coculture of vascular smooth muscle cells (SMCs) with vascular endothelial cell (ECs) 14 15 , or HASMCs with human umbilical vein endothelial cells (HUVECs) 16 .…”

“…Recently, hASCs have demonstrated the potential for differentiation into multiple cell lineages, including osteocytes, chondrocytes, smooth muscle cells and endothelial cells 7 8 9 10 11 12 13 14 15 16 17 18 19 20 . In addition, hASCs are easier to obtain and exhibit lower donor-site morbidity.…”

Bioreactors for tissue engineering: An update

“…For vascular tissue engineering, bioreactor technology is still under-explored but bioreactors that provide adequate perfusion and fluid flow has shown to provide an optimal environment for adequate tissue formation [43]. Future work will need to specify the optimal cell source used to create an optimal tissue engineered vascular construct [75]. With the rapid expansion in skin tissue engineering, multiple bioreactors have been utilised for in vitro skin generation.…”

3D Tissue-Engineered Vascular Drug Screening Platforms: Promise and Considerations

“…In this study, ASCs-derived smooth muscle cells were seeded into electrospun polycaprolactone-gelatin scaffolds and maturation was induced in a pulsatile bioreactor followed by seeding with endothelial cells differentiated from ASCs (63). Another study compared the proteomic profiles of normal arterial walls with tissue engineered blood vessels developed by ASCs-derived smooth muscle cells seeded into polyglycolic acid scaffolds and conditioned in a pulsatile bioreactor (62). The study identified 38 differentially expressed proteins between normal vessels and engineered grafts, the majority of which were cytoskeletal and actin-related proteins, indicating altered cytoskeletal integrity in ASCs engineered tissues (62).…”