Direct functionalization of cell-adhesion promoters to hydrogel microparticles synthesized by stop-flow lithography

초록

Polyethylene glycol (PEG) hydrogel microparticles generated via stop-flow lithography can be utilized for efficient microparticle-based cell culture processes because of their high biocompatibility, the molecular diffusion capability in the gel structure, and the tunability of their shape and size. However, the typical functionalization process of PEG microparticles with cell-adhesion promoters has inevitable limitations, requiring additional linker molecules and the preconjugation of linkers to cell-adhesion promoters and microparticles. In this study, a simple and direct cell-adhesion promoter functionalization process of the PEG microparticles is presented by use of aza-Michael reaction between remnant unreacted acrylate groups in particles and amine groups in cell-adhesion promoters. On the basis of proposed process, particles are directly conjugated with poly-l-lysine (PLL), a typical cell-adhesion promoter that can electrostatically interact with cellular membranes, in a controllable manner. We demonstrate enhanced cell-adhesion capabilities of the particles along with the increased amount of conjugated PLL in the particles. Furthermore, to validate extended applicability, the particles are directly conjugated with Gly-Arg-Gly-Asp-Ser (GRGDS) peptides, in which RGD sequence is involved in the cell-adhesion behavior of extracellular matrix proteins, including fibronectin. The introduced GRGDS peptides increase the cell-adhesion capacity of the microparticles binding to integrin proteins in cellular membranes.

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