A Sessile Drop Method for Facile and Robust Spheroid Cultures
A Sessile Drop Method for Facile and Robust Spheroid Cultures
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DOI：
10.1002/admi.202100972
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发表时间：
2021-09
影响因子：
5.4
通讯作者：
Bangyong Sun;Yi Zhao;Q. Zhao;Gang Li
中科院分区：
文献类型：
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作者：
Bangyong Sun;Yi Zhao;Q. Zhao;Gang Li
在这项工作中，提出了一种创新的固着滴（SD）策略，用于简便、稳健和长期的细胞球体培养。与传统的悬滴 (HD) 方法不同，该策略采用超疏水穿孔微孔板 (SHPMP) 在空间上限制细胞悬浮液滴阵列，用于 3D 球体培养。受益于细胞悬浮液滴的向上定位和微孔表面的不可润湿性的结合，SD培养平台不仅保持了HD方法的技术简单性和高氧气利用率，而且还提供了高机械稳定性和易于培养基更换，从而能够以高通量方式实现稳健的长期球体培养。在概念验证应用中，通过使用 SHPMP 生成 384 个均匀的 MHCC97H 细胞球体，证明了 SD 方法的便利性和简单性。为了验证该方法的实用性，进一步证明该方法制备的球体可用于药物测试。 SD平台具有液体处理方便、试剂/样品消耗量低、液滴稳定性高、供氧充足以及良好的原位监测兼容性等优点，有利于细胞球体的长期高通量培养，为组织工程、再生医学和药物筛选提供了有力的工具。
In this work, an innovative sessile drop (SD) strategy for facile, robust, and long‐term cell spheroid cultures is proposed. Unlike the conventional hanging drop (HD) method, this strategy employs a superhydrophobic perforated microwell plate (SHPMP) to spatially confine arrays of cell suspension droplets for 3D spheroid cultures. Benefitting from a combination of upward positioning of cell suspension droplets and non‐wettability of the microwell surface, the SD culture platform not only maintains the technical simplicity and high oxygen availability of the HD method, but also provides high mechanical stability and ease of medium exchange, thus enabling a robust long‐term spheroid culture in a high‐throughput manner. In a proof of concept application, the convenience and simplicity of the SD approach is demonstrated by producing 384 uniform spheroids of MHCC97H cell with SHPMP. To validate the utility of this method, it is further demonstrated that spheroids fabricated by this method can be used for drug testing. With convenient liquid handling, low reagent/sample consumption, high droplet stability, adequate oxygen supply, and good in situ monitoring compatibility, the SD platform is beneficial for long‐term cell spheroid culture in a high throughput manner and provides a powerful tool for tissue engineering, regenerative medicine, and drug screening.