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用于收获细胞和细胞片的温敏性微凝胶薄膜。

Thermoresponsive microgel films for harvesting cells and cell sheets.

机构信息

State Key Laboratory of Heavy Oil Processing and the Centre for Bioengineering and Biotechnology, China University of Petroleum , East China, Qingdao, 266555, China.

出版信息

Biomacromolecules. 2013 Oct 14;14(10):3615-25. doi: 10.1021/bm4009765. Epub 2013 Sep 13.

Abstract

This work reports the formation of thermoresponsive poly(N-isopropylacrylamide-co-styrene) (PNIPAAmSt) microgel films and their use for cell growth and detachment via temperature stimuli. Thermoresponsive surface films can be conveniently produced by spin-coating or drop-coating of PNIPAAmSt microgel dispersions onto substrates such as glass coverslips, cell culture plates, and flasks, making this technique widely accessible. The thickness, stability, and reversibility of the PNIPAAmSt films coated on silicon wafers with respect to temperature switching were examined by spectroscopic ellipsometry (SE) and atomic force microscopy (AFM). The results unraveled the direct link between thermoreversibility and changes in film thickness and surface morphology, showing reversible hydration and dehydration. Under different coating conditions, well-packed microgel monolayers could be utilized for effective cell recovery and harvesting. Furthermore, cell adhesion and detachment processes were reversible and there was no sign of loss of cell viability during repeated surface attachment, growth, and detachment, showing a mild interaction between cells and thermoresponsive surface. More importantly, there was little deterioration of the packing of the thermoresponsive films or any major loss of microgel particles during reuse, indicating their robustness. These PNIPAAmSt microgel films thus open up a convenient interfacial platform for cell and cell sheet harvesting while avoiding the damage of enzymatic cleavage.

摘要

这项工作报道了温敏性聚(N-异丙基丙烯酰胺-共-苯乙烯)(PNIPAAmSt)微凝胶薄膜的形成及其在细胞生长和通过温度刺激的细胞脱落方面的应用。温敏表面薄膜可以通过将 PNIPAAmSt 微凝胶分散体旋涂或滴涂到例如玻璃盖玻片、细胞培养板和培养瓶等基底上来方便地制备,使得这种技术广泛可用。通过光谱椭圆术(SE)和原子力显微镜(AFM)检查了涂覆在硅晶片上的 PNIPAAmSt 薄膜在温度切换时的厚度、稳定性和可逆性。结果揭示了热可逆性与薄膜厚度和表面形貌变化之间的直接联系,表明了可逆的水合和去水合作用。在不同的涂层条件下,可以利用紧密堆积的微凝胶单层来有效回收和收获细胞。此外,细胞附着和脱落过程是可逆的,并且在重复的表面附着、生长和脱落过程中没有细胞活力损失的迹象,表明细胞与温敏表面之间存在温和的相互作用。更重要的是,在重复使用过程中,温敏薄膜的包装几乎没有恶化,也没有微凝胶颗粒的大量损失,表明其坚固耐用。因此,这些 PNIPAAmSt 微凝胶薄膜为细胞和细胞片的收获提供了一个方便的界面平台,同时避免了酶切造成的损伤。

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