A pillar/perfusion plate enhances cell growth, reproducibility, throughput, and user friendliness in dynamic 3D cell culture.
A pillar/perfusion plate enhances cell growth, reproducibility, throughput, and user friendliness in dynamic 3D cell culture.
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柱/灌注板可增强动态 3D 细胞培养中的细胞生长、重现性、通量和用户友好性。
DOI：
10.1101/2023.02.16.528892
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Lee,Moo-Yeal
中科院分区：
文献类型：
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作者：
ReddyLekkala,VinodKumar;Kang,Soo-Yeon;Liu,Jiafeng;Shrestha,Sunil;Acharya,Prabha;Joshi,Pranav;Zolfaghar,Mona;Lee,Minseong;Jamdagneya,Paarth;Pagnis,Sohan;Kundi,Arham;Kabbur,Samarth;Kim,UngTae;Yang,Yong;Lee,Moo-Yeal
静态三维（3D）细胞培养已经在超低附着孔板、悬滴板和具有水凝胶或磁性纳米颗粒的微量滴定孔板中得到证实。尽管静态培养的3D细胞简单、可重复且相对便宜，因此有可能用于高通量筛选，但由于营养和氧气扩散以及废物清除有限，静态培养的3D细胞通常会出现坏死核心，并且具有有限的体内样组织结构。在这里，我们通过开发柱/灌注板平台并展示高通量动态3D细胞培养来克服这些挑战。通过简单的包埋和包裹将细胞球负载在水凝胶柱板上，并在数字摇杆上的灌注板中进行动态培养。与传统的微流体装置不同，在灌注威尔斯孔内保持快速流速，并且柱板与灌注板分离以用于基于细胞的测定。它与普通实验室设备兼容，并允许细胞培养，测试，染色和原位成像。柱/灌注板通过在动态3D细胞培养中的快速扩散、再现性、测定通量和用户友好性来增强细胞生长。
Static three-dimensional (3D) cell culture has been demonstrated in ultralow attachment well plates, hanging droplet plates, and microtiter well plates with hydrogels or magnetic nanoparticles. Although it is simple, reproducible, and relatively inexpensive, thus potentially used for high-throughput screening, statically cultured 3D cells often suffer from a necrotic core due to limited nutrient and oxygen diffusion and waste removal and have a limitedin vivo-like tissue structure. Here, we overcome these challenges by developing a pillar/perfusion plate platform and demonstrating high-throughput, dynamic 3D cell culture. Cell spheroids were loaded on the pillar plate with hydrogel by simple sandwiching and encapsulation and cultured dynamically in the perfusion plate on a digital rocker. Unlike traditional microfluidic devices, fast flow velocity was maintained within perfusion wells and the pillar plate was separated from the perfusion plate for cell-based assays. It was compatible with common lab equipment and allowed cell culture, testing, staining, and imagingin situ. The pillar/perfusion plate enhanced cell growth by rapid diffusion, reproducibility, assay throughput, and user friendliness in a dynamic 3D cell culture.