Development of non-adherent cell-enclosing domes with enzymatically cross-linked hydrogel shell
Development of non-adherent cell-enclosing domes with enzymatically cross-linked hydrogel shell
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DOI：
10.1088/1758-5090/ac95ce
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发表时间：
2023-01-01
期刊：
影响因子：
9
通讯作者：
Sakai, Shinji
中科院分区：
文献类型：
--
作者：
Kazama, Ryotaro;Sato, Ryuta;Sakai, Shinji
非贴壁细胞，如造血细胞和淋巴细胞，是医学和生物学领域的重要研究课题。因此，一种能够处理溶液中的非粘连细胞的系统将是有用的，该系统能够以与在介质交换、接触化学品、洗涤和成像应用中的染色过程中粘连细胞相同的方式处理非粘连细胞。在这里，我们报道了一种‘细胞穹顶’平台，在该平台上，未贴壁的细胞可以被封闭并生长在直径约1 mm、高度约270微米的空腔中。穹顶由90微米厚的海藻酸盐水凝胶壳组成。通过辣根过氧化物酶介导的交联，在玻璃板上形成细胞穹顶。人白血病细胞系K562细胞包被在细胞穹顶内，稳定29天，每隔2~3天换液一次。封闭的细胞在腔内生长，并用周围培养液提供的试剂进行染色和分化。此外，填充空洞(3D微环境)的K562细胞比2D培养的K562细胞更低氧，对丝裂霉素C具有更高的抵抗力。这些发现表明，“细胞穹顶”可能是一种很有前途的工具，可以方便地培养和评估非贴壁细胞。
Non-adherent cells, such as hematopoietic cells and lymphocytes, are important research subjects in medical and biological fields. Therefore, a system that enables the handling of non-adherent cells in solutions in the same manner as that of adhering cells during medium exchange, exposure to chemicals, washing, and staining in imaging applications would be useful. Here, we report a 'Cell Dome' platform in which non-adherent cells can be enclosed and grown in the cavities of about 1 mm diameter and 270 mu m height. The domes consist of an alginate-based hydrogel shell of 90 mu m thickness. Cell Domes were formed on glass plates by horseradish peroxidase-mediated cross-linking. Human leukaemia cell line K562 cells enclosed in Cell Domes were stable for 29 days with every 2-3 days of medium change. The enclosed cells grew in the cavities and were stained and differentiated with reagents supplied from the surrounding medium. Additionally, K562 cells that filled the cavities (a 3D microenvironment) were more hypoxic and highly resistant to mitomycin C than those cultured in 2D. These findings demonstrate that the 'Cell Dome' may be a promising tool for conveniently culturing and evaluating non-adherent cells.