Application of DNA Quadruplex Hydrogels Prepared from Polyethylene Glycol-Oligodeoxynucleotide Conjugates to Cell Culture Media
Application of DNA Quadruplex Hydrogels Prepared from Polyethylene Glycol-Oligodeoxynucleotide Conjugates to Cell Culture Media
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DOI：
10.3390/polym11101607
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发表时间：
2019-10
期刊：
影响因子：
5
通讯作者：
S. Tanaka;Shinsuke Yukami;Yuhei Hachiro;Y. Ohya;A. Kuzuya
中科院分区：
文献类型：
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作者：
S. Tanaka;Shinsuke Yukami;Yuhei Hachiro;Y. Ohya;A. Kuzuya
应用Na+响应的DNA四链体水凝胶，利用G-四链体作为交联点的聚（乙二醇）（PEG）网络作为细胞培养基质，已被检查。采用改进的高效液相法（HELP）合成了聚乙二醇（PEG）-脱氧鸟苷（dG 4）偶联物，并将其作为大分子单体。当与等体积的细胞培养基混合时，PEG-ODN的溶液由于在Na+存在下dG 4片段形成G-四链体而变成刚性水凝胶（G-四链体水凝胶）。PEG-ODN本身不显示细胞毒性，并且所得水凝胶在细胞培养条件下足够稳定。然而，在G-四链体水凝胶中培养的L929成纤维细胞保持球形一周，仍然存活，没有增殖。细胞逐渐沉降通过凝胶一天一天，可能是由于G-四链体形成的可逆性和由此产生的大分子单体的缓慢重排。一旦它们到达底部玻璃表面，细胞就开始扩散和增殖。
Application of Na+-responsive DNA quadruplex hydrogels, which utilize G-quadruplexes as crosslinking points of poly(ethylene glycol) (PEG) network as cell culture substrate, has been examined. PEG-oligodeoxynucleotide (ODN) conjugate, in which four deoxyguanosine (dG4) residues are tethered to both ends of PEG, was prepared by modified high-efficiency liquid phase (HELP) synthesis of oligonucleotides and used as the macromonomer. When mixed with equal volume of cell culture media, the solution of PEG-ODN turned into stiff hydrogel (G-quadruplex hydrogel) as the result of G-quadruplex formation by the dG4 segments in the presence of Na+. PEG-ODN itself did not show cytotoxicity and the resulting hydrogel was stable enough under cell culture conditions. However, L929 fibroblast cells cultured in G-quadruplex hydrogel remained spherical for a week, yet alive, without proliferation. The cells gradually sedimented through the gel day by day, probably due to the reversible nature of G-quadruplex formation and the resulting slow rearrangement of the macromonomers. Once they reached the bottom glass surface, the cells started to spread and proliferate.