A DNA/Poly-(L-lysine) Hydrogel with Long Shelf-Time for 3D Cell Culture.
Jianpu TangJing WangJunhan OuZhen CuiChi YaoDayong YangPublished in: Small methods (2024)
Deoxyribonucleic acid (DNA)-based hydrogels are emerging as promising functional materials for biomedical applications. However, the shelf-time of DNA hydrogels in biological media is severely shortened by nucleases, which limit the application of DNA hydrogels. Herein, a DNA hydrogel with long shelf-time is reported for 3D cell culture. Poly-(L-lysine) (PLL) is introduced as both a cross-linker and a protectant. The electrostatic interaction between PLL and DNA drove the formation of hydrogel. PLL coating on DNA increased the steric hindrance between DNA and nucleases, thus weakening the digestion of nucleases toward phosphodiester bond. As a result, the shelf-time of DNA/PLL hydrogel for 3D cell culture is extended from generally 1 day to longer than 15 days, which has not been achieved previously. Notably, poly-AS1411-aptamers are integrated to DNA/PLL hydrogels for anchoring U87 cells, and the cell encapsulation efficiency of the DNA/PLL hydrogels with aptamer is 4-time higher than that of the hydrogels without aptamer. DNA/PLL hydrogel provided a favorable microenvironment to support the proliferation of cells, which formed cell spheroid in 15 days. This protective coating strategy solves the long-standing problem on the shelf-time of DNA hydrogel, and is envisioned to promote the development of DNA hydrogel in more biomedical applications.
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