Surface-tension driven open microfluidic platform for hanging droplet culture.
Surface-tension driven open microfluidic platform for hanging droplet culture.
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DOI：
10.1039/c5lc01353d
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发表时间：
2016-01-21
期刊：
影响因子：
6.1
通讯作者：
Theberge AB
中科院分区：
文献类型：
--
作者：
de Groot TE;Veserat KS;Berthier E;Beebe DJ;Theberge AB
用于三维组织培养的悬滴技术已经在生物实验室中使用了几十年，其核心技术相对保持不变。最近，微尺度方法扩展了悬滴方法的能力，使其更加用户友好。我们提出了一种自发驱动的开放悬挂液滴培养平台，以解决当前平台的许多局限性。我们的平台利用两个相互连接的悬滴威尔斯孔，一个较大的孔用于培养细胞，一个较小的孔用于通过移液管进行用户界面。双孔系统在流体交换期间在培养孔中产生较低的剪切应力，使得剪切敏感性或非粘附性细胞能够在液滴中培养。在液滴中进行流体交换的能力使得能够在不破坏培养物的情况下进行长期培养、处理和表征。该平台的开放式孔格式用于进行时间依赖性共培养，从而实现具有悬浮生长的骨组织支架和细胞的培养配置。该系统的开放性质允许在实验过程中直接添加或移除组织，这在其他微流体或悬滴培养平台中是不切实际的操作。
The hanging droplet technique for three-dimensional tissue culture has been used for decades in biology labs, with the core technology remaining relatively unchanged. Recently microscale approaches have expanded the capabilities of the hanging droplet method, making it more user-friendly. We present a spontaneously driven, open hanging droplet culture platform to address many limitations of current platforms. Our platform makes use of two interconnected hanging droplet wells, a larger well where cells are cultured and a smaller well for user interface via a pipette. The two-well system results in lower shear stress in the culture well during fluid exchange, enabling shear sensitive or non-adherent cells to be cultured in a droplet. The ability to perform fluid exchanges in-droplet enables long-term culture, treatment, and characterization without disruption of the culture. The open well format of the platform was utilized to perform time-dependent coculture, enabling culture configurations with bone tissue scaffolds and cells grown in suspension. The open nature of the system allowed the direct addition or removal of tissue over the course of an experiment, manipulations that would be impractical in other microfluidic or hanging droplet culture platforms.