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具有可调节组成的多糖基水凝胶作为3D细胞培养系统。

Polysaccharide-based hydrogels with tunable composition as 3D cell culture systems.

作者信息

Gentilini Roberta, Munarin Fabiola, Bloise Nora, Secchi Eleonora, Visai Livia, Tanzi Maria Cristina, Petrini Paola

机构信息

1 Department of Chemistry, Materials and Chemical Engineering, Unità di Ricerca Consorzio INSTM, Politecnico di Milano, Milan, Italy.

2 Department of Electronics, Information and Bioengineering, Politecnico di Milano, Milan, Italy.

出版信息

Int J Artif Organs. 2018 Apr;41(4):213-222. doi: 10.5301/ijao.5000667. Epub 2018 Feb 19.

Abstract

BACKGROUND

To date, cell cultures have been created either on 2-dimensional (2D) polystyrene surfaces or in 3-dimensional (3D) systems, which do not offer a controlled chemical composition, and which lack the soft environment encountered in vivo and the chemical stimuli that promote cell proliferation and allow complex cellular behavior. In this study, pectin-based hydrogels were developed and are proposed as versatile cell culture systems.

METHODS

Pectin-based hydrogels were produced by internally crosslinking pectin with calcium carbonate at different initial pH, aiming to control crosslinking kinetics and degree. Additionally, glucose and glutamine were added as additives, and their effects on the viscoelastic properties of the hydrogels and on cell viability were investigated.

RESULTS

Pectin hydrogels showed in high cell viability and shear-thinning behavior. Independently of hydrogel composition, an initial swelling was observed, followed by a low percentage of weight variation and a steady-state stage. The addition of glucose and glutamine to pectin-based hydrogels rendered higher cell viability up to 90%-98% after 1 hour of incubation, and these hydrogels were maintained for up to 7 days of culture, yet no effect on viscoelastic properties was detected.

CONCLUSIONS

Pectin-based hydrogels that offer tunable composition were developed successfully. They are envisioned as synthetic extracellular matrix (ECM) either to study complex cellular behaviors or to be applied as tissue engineering substitutes.

摘要

背景

迄今为止,细胞培养是在二维(2D)聚苯乙烯表面或三维(3D)系统中进行的,这些系统无法提供可控的化学成分,缺乏体内所具有的柔软环境以及促进细胞增殖并允许复杂细胞行为的化学刺激。在本研究中,开发了基于果胶的水凝胶,并将其作为通用的细胞培养系统。

方法

通过在不同初始pH值下将果胶与碳酸钙进行内部交联来制备基于果胶的水凝胶,旨在控制交联动力学和交联程度。此外,添加葡萄糖和谷氨酰胺作为添加剂,并研究它们对水凝胶粘弹性特性和细胞活力的影响。

结果

果胶水凝胶表现出高细胞活力和剪切变稀行为。无论水凝胶组成如何,均观察到初始膨胀,随后重量变化百分比低且进入稳态阶段。向基于果胶的水凝胶中添加葡萄糖和谷氨酰胺,在孵育1小时后细胞活力提高至90%-98%,并且这些水凝胶可维持长达7天的培养,但未检测到对粘弹性特性有影响。

结论

成功开发了具有可调组成的基于果胶的水凝胶。它们被设想为合成细胞外基质(ECM),用于研究复杂的细胞行为或用作组织工程替代物。

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