Inter-donor variability of extracellular matrix production in long-term cultures of human fibroblasts

Abstract

Several tissue engineering approaches are based on the ability of mesenchymal cells to endogenously synthetize an extracellular matrix (ECM) in vitro, which can be seen as a form of biomaterial....

Development and characterization of biological sutures made of cell-assembled extracellular matrix

“…Human and ovine biological materials synthesized in the laboratory by normal dermal fibroblasts in culture has been developed [19][20][21][22][23][24]. This material, called Cell-Assembled extracellular Matrix (CAM), is produced in large culture flasks as a sheet, which can be cut into ribbons that can be assembled in biological textiles [21,22,25].…”

Successive Protein Extraction Using Hydroxylamine to Increase the Depth of Proteome Coverage in Fresh, Forensic, and Archaeological Bones

“…Key advantages of the use of this reagent is that it has a significantly lower toxicity compared to CNBr, the digestion step can be easily incorporated into a classical proteomic preparation protocol (as shown in Figure 1B,C, thus reducing sample loss), and it has been shown to be more reproducible than CNBr in proteomic studies. 29,30 Here, we propose the use of a staggered extraction protocol, illustrated in Figure 1B, wherein a chemical digestion is used in order to solubilize the typically inaccessible part of the collagen matrix. Through the addition of an extra extraction step on a fraction of the sample that is typically discarded, the collagen extraction yield is significantly enhanced, with an increase in the signal-to-noise ratio due to the increased peptide signal of peptides corresponding to the core extracellular matrix proteins in bone.…”

“…We also propose a protocol wherein hydroxylamine extraction is applied directly on the demineralized bone, targeting collagen extraction, as shown in Figure C. By adapting a protocol for archeological bone proteomics with the inclusion of a hydroxylamine digestion step, we evaluate the added value of the additional extraction applied to several types of mineralized tissues, including both modern (fresh bovine bone, human anatomical specimens) and archeological bones and teeth. , …”

“…By adapting a protocol for archeological bone proteomics with the inclusion of a hydroxylamine digestion step, we evaluate the added value of the additional extraction applied to several types of mineralized tissues, including both modern (fresh bovine bone, human anatomical specimens) and archeological bones and teeth. 29,30 ■ MATERIALS AND METHODS Samples. Fresh bovine bone samples, human anatomical specimens (forensic), and human archeological samples of different preservation states were used in this study.…”

“…This imide structure then comes under nucleophilic attack by hydroxylamine, which results in the cleavage of the N–C peptide bond. Key advantages of the use of this reagent is that it has a significantly lower toxicity compared to CNBr, the digestion step can be easily incorporated into a classical proteomic preparation protocol (as shown in Figure B,C, thus reducing sample loss), and it has been shown to be more reproducible than CNBr in proteomic studies. , …”