Abstract
Hydrogels have strong application prospects for drug delivery, tissue engineering and cell therapy because of their excellent biocompatibility and abundant availability as scaffolds for drugs and cells. In this study, we created hybrid hydrogels based on a genetically modified tax interactive protein-1 (TIP1) by introducing two or four cysteine residues in the primary structure of TIP1. The introduced cysteine residues were crosslinked with a four-armed poly (ethylene glycol) having their arm ends capped with maleimide residues (4-armed-PEG-Mal) to form hydrogels. In one form of the genetically modification, we incorporated a peptide sequence ‘GRGDSP’ to introduce bioactivity to the protein, and the resultant hydrogel could provide an excellent environment for a three dimensional cell culture of AD293 cells. The AD293 cells continued to divide and displayed a polyhedron or spindle-shape during the 3-day culture period. Besides, AD293 cells could be easily separated from the cell-gel constructs for future large-scale culture after being cultured for 3 days and treating hydrogel with trypsinase. This work significantly expands the toolbox of recombinant proteins for hydrogel formation, and we believe that our hydrogel will be of considerable interest to those working in cell therapy and controlled drug delivery.
Highlights
By mimicking the biochemical and mechanical properties of native tissue, hydrogels possess hydrated networks of bioactive components [1], such as drugs, proteins, sugars and even cells, leading to contributions to the fields of controllable drug delivery [2,3], tissue engineering [4,5], cell culture [6,7], and others
We investigated whether the tax interactive protein-1 (TIP1) 2C gel or the TIP1 2C RGD gel was suitable for 3D cell culture
We have successfully produced in situ gelating hybrid hydrogels composed of genetically modified recombinant proteins and 4-armed-PEG-Mal
Summary
By mimicking the biochemical and mechanical properties of native tissue, hydrogels possess hydrated networks of bioactive components [1], such as drugs, proteins, sugars and even cells, leading to contributions to the fields of controllable drug delivery [2,3], tissue engineering [4,5], cell culture [6,7], and others. Cells are known to receive and respond to signals from the crosstalk of extrinsic complexes by proteins, one of the essential building blocks [8]. Protein-based hydrogels can provide excellent environments for the cells, and respond to external stimuli. More and more researchers have focused on the promising applications of protein-based hydrogels, especially in the analytic detection [9,10] and three-dimensional cell culture [11,12]. Cells exhibit expansion and adhesion in gels by integrating adhesive peptide modules such as RGDS and REDV [14]
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