Lectin-mediated reversible immobilization of human cells into a glycosylated macroporous protein hydrogel as a cell culture matrix

Abstract

3D cell culture is a helpful approach to study cell-cell interaction in a native-like environment, but is often limited due the challenge of retrieving cells from the material. In this study, we present the use of recombinant lectin B, a sugar-binding protein with four binding cavities, to enable reversible cell integration into a macroporous protein hydrogel matrix. By functionalizing hydrogel precursors with saccharose, lectin B can both bind to sugar moieties on the cellular surface as well as to the modified hydrogel network. Confocal microscopy and flow cytometry analysis revealed cells to be integrated into the network and to adhere and proliferate. Furthermore, the specificity and reversibility was investigated by using a recombinantly produced yellow fluorescent - lectin B fusion protein and a variety of sugars with diverging affinities for lectin B at different concentrations and elution times. Cells could be eluted within minutes by addition of L-fucose to the cell-loaded hydrogels to make cells available for further analysis.

Robust antigen-specific tuning of the nanoscale barrier properties of biogels using matrix-associating IgG and IgM antibodies

“…Indeed, steric trapping of micro-and nanoparticulates in Matrigel® and other highly glycosylated biologically derived hydrogels is well established [12][13][14]. Consequently, to tune the barrier properties of biogels, the predominant strategies are to either (i) alter the pore sizes of the gel matrix and impede transit through changing steric hindrance [2,15] or (ii) conjugate specific motifs to the gel itself that in turn bind foreign species [16][17][18][19][20]. Both approaches involve substantial drawbacks.…”

“…light or enzymes, respectively) to polymerize or degrade matrix constituents [20][21][22][23][24][25], only facilitate selectivity based on size and are generally irreversible. Alternatively, covalent crosslinking of antigen-specific molecules to the matrix [16][17][18][19]25] is cumbersome in practice, is limited in the number of species that can be simultaneously immobilized, and may not efficiently immobilize diffusive species [26].…”

A Cerberus‐Inspired Anti‐Infective Multicomponent Gatekeeper Hydrogel against Infections with the Emerging “Superbug” Yeast Candida auris

“…[ 13 ] AMPs have been loaded into hydrogels for topical applications [ 16,17 ] on the wound surface at the infection site and to achieve first line of defense against fatal wound infections, for example, for applications in intensive care units and after surgery of contaminated patients. The mannose‐specific lectin B (LecB) as a tetrameric protein providing four binding sites toward sugar residues on the surface of bacteria and higher cells [ 18,19 ] has already been used to bind and immobilize carbapenem‐resistant Pseudomonas aeruginosa within a hydrogel matrix that were subsequently eradicated by the respective AMPs. [ 15 ]…”

Gelatin/dextran‐based hydrogel cross‐linked by Diels–Alder click chemistry: the swelling and potassium diclofenac releasing

“…Furthermore, hydrogels could be obtained from a wide variety of materials, since a natural source to synthetic ones, being applied in food, agronomy and biomedicine (Hoffman, 2012). The applications in the biomedicine field are focus on scaffolds for tissue engineering (Bai et al, 2018), cellular immobilization (Bodenberger et al, 2017) and drug delivery systems (Pulickal et al, 2018).…”