Abstract:
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 limited in 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 imaging in situ. The pillar/perfusion plate enhanced cell growth by rapid diffusion, reproducibility, assay throughput, and user friendliness in a dynamic 3D cell culture.
摘要:
静态三维(3D)细胞培养已在超低附着孔板中得到证明,悬挂液滴板,和具有水凝胶或磁性纳米颗粒的微量滴定孔板。虽然很简单,可重复,相对便宜,因此有可能用于高通量筛选,静态培养的3D细胞通常由于有限的营养和氧扩散以及废物去除而遭受坏死核,并且具有有限的体内样组织结构。这里,我们通过开发支柱/灌注板平台和展示高通量来克服这些挑战,动态3D细胞培养。通过简单的夹心和封装将细胞球体加载到具有水凝胶的柱板上,并在数字摇杆上的灌注板中动态培养。与传统的微流体装置不同,在灌注孔中保持快速流速,并将柱板与灌注板分离以进行基于细胞的测定.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/灌注板通过快速扩散增强细胞生长,再现性,测定通量,以及动态3D细胞培养中的用户友好性。