Article ; Online: Lectin-mediated reversible immobilization of human cells into a glycosylated macroporous protein hydrogel as a cell culture matrix.
2017 Volume 7, Issue 1, Page(s) 6151
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 ... ...
| 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. |
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| MeSH term(s) | A549 Cells ; Cell Adhesion ; Cell Culture Techniques/methods ; Cell Proliferation ; Flow Cytometry ; Glycosylation ; Humans ; Hydrogels/chemistry ; Microscopy, Confocal ; Pokeweed Mitogens/chemistry ; Pokeweed Mitogens/metabolism ; Porosity ; Recombinant Proteins/chemistry ; Recombinant Proteins/metabolism ; Sucrose/metabolism |
| Chemical Substances | Hydrogels ; Pokeweed Mitogens ; Recombinant Proteins ; lectin B, Phytolacca americana ; Sucrose (57-50-1) |
| Language | English |
| Publishing date | 2017-07-21 |
| Publishing country | England |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ZDB-ID | 2615211-3 |
| ISSN | 2045-2322 ; 2045-2322 |
| ISSN (online) | 2045-2322 |
| ISSN | 2045-2322 |
| DOI | 10.1038/s41598-017-06240-w |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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