Ogiwara Naoto, Nakano Takenobu, Masuda Tsukuru, Kushiro Keiichiro, Takai Madoka
Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
Corporate R&D Division, Toyo Ink SC Holdings Co., Ltd., 5-6-7 Chiyoda, Sakado-city, Saitama, 350-0214, Japan.
Macromol Biosci. 2023 May;23(5):e2200486. doi: 10.1002/mabi.202200486. Epub 2023 Mar 12.
3D structured cells have great drug screening potential because they mimic in vivo tissues better than 2D cultured cells. In this study, multi-block copolymers composed of poly(2-methoxyethyl acrylate) (PMEA) and polyethylene glycol (PEG) are developed as a new kind of biocompatible polymers. PEG imparts non-cell adhesion while PMEA acts as an anchoring segment to prepare the polymer coating surface. The multi-block copolymers show higher stability in water than PMEA. A specific micro-sized swelling structure composed of a PEG chain is observed in the multi-block copolymer film in water. A single NIH3T3-3-4 spheroid is formed in 3 h on the surface of the multi-block copolymers with 8.4 wt% PEG. However, at a PEG content of 0.7 wt%, spheroid formed after 4 days. The adenosine triphosphate (ATP) activity of cells and the internal necrotic state of the spheroid change depending on PEG loading in the multi-block copolymers. As the formation rate of cell spheroid on low-PEG-ratio multi-block copolymers is slow, internal necrosis of cell spheroid is less likely to occur. Consequently, the cell spheroid formation rate by changing the PEG chain content in multi-block copolymers is successfully controlled. These unique surfaces are suggested to be useful for 3D cell culture.
3D结构化细胞具有巨大的药物筛选潜力，因为它们比二维培养细胞能更好地模拟体内组织。在本研究中，由聚（丙烯酸2-甲氧基乙酯）（PMEA）和聚乙二醇（PEG）组成的多嵌段共聚物被开发为一种新型的生物相容性聚合物。PEG赋予非细胞粘附性，而PMEA作为锚定链段来制备聚合物涂层表面。多嵌段共聚物在水中比PMEA具有更高的稳定性。在水中的多嵌段共聚物薄膜中观察到由PEG链组成的特定微尺寸溶胀结构。在含有8.4 wt% PEG的多嵌段共聚物表面，单个NIH3T3-3-4球体在3小时内形成。然而，当PEG含量为0.7 wt%时，4天后形成球体。细胞的三磷酸腺苷（ATP）活性和球体的内部坏死状态随多嵌段共聚物中PEG负载量的变化而变化。由于低PEG比例多嵌段共聚物上细胞球体的形成速率较慢，细胞球体内部坏死的可能性较小。因此，通过改变多嵌段共聚物中PEG链的含量成功地控制了细胞球体的形成速率。这些独特的表面被认为对3D细胞培养有用。
