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1.
Bioconjug Chem ; 25(9): 1626-31, 2014 Sep 17.
Article in English | MEDLINE | ID: mdl-25133293

ABSTRACT

In the current study, synthetic hydrogels containing metabolically engineered glycoproteins of mammalian cells were prepared for the first time and selectin-mediated cell adhesion on the hydrogel was demonstrated. A culture of HL-60 cells was supplemented with an appropriate volume of aqueous solution of N-methacryloyl mannosamine (ManMA) to give a final concentration of 5 mM. The cells were then incubated for 3 days to deliver methacryloyl groups to the glycoproteins of the cells. A transparent hydrogel was formed via redox radical polymerization of methacryloyl functionalized glycoproteins with 2-methacryloyloxyethyl phosphorylcholine and a cross-linker. Conjugation of the glycoproteins into the hydrogel was determined using Coomassie brilliant blue (CBB) and periodic acid-Schiff (PAS) staining. The surface density of P-selectin glycoprotein ligand-1 (PSGL-1) on the hydrogels was also detected using gold-colloid-labeled immunoassay. Finally, selectin-mediated cell adhesion on hydrogels containing glycoproteins was demonstrated. Selectin-mediated cell adhesion is considered an essential step in the progression of various diseases; therefore, hydrogels having glycoproteins could be useful in therapeutic and diagnostic applications.


Subject(s)
Glycoproteins/chemistry , Glycoproteins/pharmacology , Hydrogels/chemistry , Hydrogels/chemical synthesis , Metabolic Engineering , Oligosaccharides/metabolism , Cell Adhesion/drug effects , Glycoproteins/metabolism , HL-60 Cells , Hexosamines/chemistry , Human Umbilical Vein Endothelial Cells/cytology , Human Umbilical Vein Endothelial Cells/drug effects , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Hydrogels/metabolism , P-Selectin/metabolism
2.
Chem Commun (Camb) ; 49(71): 7824-6, 2013 Sep 14.
Article in English | MEDLINE | ID: mdl-23884331

ABSTRACT

In order to introduce alternative functions into mammalian cells and control them under ambient conditions, poly(N-isopropyl acrylamide) (PNIPAM) was immobilized on the cell surface. Cellular aggregation could be regulated by temperature change. In addition, separation of PNIPAM-conjugated glycoproteins was successfully performed.


Subject(s)
Acrylic Resins/chemistry , HL-60 Cells , Humans , Membrane Glycoproteins/chemistry , Membrane Glycoproteins/metabolism , Temperature
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