Institute of Physiologically Active Compounds of the Russian Academy of Sciences, 1 Severnij proezd, 142432 Chernogolovka, Russia.
Center of Composite Material, National University of Science and Technology "MISIS", Leninsky pr. 4, 119049 Moscow, Russia.
Molecules. 2022 Mar 24;27(7):2087. doi: 10.3390/molecules27072087.
Cell culturing methods in its classical 2D approach have limitations associated with altered cell morphology, gene expression patterns, migration, cell cycle and proliferation. Moreover, high throughput drug screening is mainly performed on 2D cell cultures which are physiologically far from proper cell functions resulting in inadequate hit-compounds which subsequently fail. A shift to 3D culturing protocols could solve issues with altered cell biochemistry and signaling which would lead to a proper recapitulation of physiological conditions in test systems. Here, we examined porous ultra-high molecular weight polyethylene (UHMWPE) as an inexpensive and robust material with varying pore sizes for cell culturing. We tested and developed culturing protocols for immortalized human neuroblastoma and primary mice hippocampal cells which resulted in high rate of cell penetration within one week of cultivation. UHMWPE was additionally functionalized with gelatin, poly-L-lysine, BSA and chitosan, resulting in increased cell penetrations of the material. We have also successfully traced GFP-tagged cells which were grown on a UHMWPE sample after one week from implantation into mice brain. Our findings highlight the importance of UHMWPE use as a 3D matrix and show new possibilities arising from the use of cheap and chemically homogeneous material for studying various types of cell-surface interactions further improving cell adhesion, viability and biocompatibility.
细胞培养方法在其经典的 2D 方法中有局限性,与改变细胞形态、基因表达模式、迁移、细胞周期和增殖有关。此外,高通量药物筛选主要在 2D 细胞培养物上进行,而 2D 细胞培养物在生理上与适当的细胞功能相差甚远,导致有效化合物不足,随后失败。转向 3D 培养方案可以解决改变细胞生物化学和信号传递的问题,从而在测试系统中正确再现生理条件。在这里,我们研究了多孔超高分子量聚乙烯(UHMWPE)作为一种具有不同孔径的廉价且坚固的材料,用于细胞培养。我们测试和开发了永生化人神经母细胞瘤和原代小鼠海马细胞的培养方案,结果在培养一周内细胞穿透率很高。UHMWPE 还通过明胶、聚-L-赖氨酸、BSA 和壳聚糖进行了功能化,从而增加了材料的细胞穿透率。我们还成功地追踪了 GFP 标记的细胞,这些细胞在植入小鼠大脑一周后在 UHMWPE 样本上生长。我们的研究结果强调了 UHMWPE 作为 3D 基质的重要性,并展示了使用廉价且化学均匀的材料进行研究各种类型的细胞表面相互作用的新可能性,进一步提高了细胞的粘附性、活力和生物相容性。