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三维细胞培养用于研究鼻息肉。

Three-dimensional cell culture for the study of nasal polyps.

机构信息

Universidade de Santo Amaro (UNISA), Programa de Pós-Graduação em Ciências da Saúde, São Paulo, SP, Brazil.

Universidade de Santo Amaro (UNISA), Programa de Pós-Graduação em Ciências da Saúde, São Paulo, SP, Brazil; Universidade Federal de São Paulo, Departamento de Otorrinolaringologia e Cirurgia de Cabeça e Pescoço, Laboratório de Pesquisa ORL, São Paulo, SP, Brazil.

出版信息

Braz J Otorhinolaryngol. 2022 Nov-Dec;88 Suppl 5(Suppl 5):S69-S74. doi: 10.1016/j.bjorl.2021.11.001. Epub 2021 Nov 26.

Abstract

OBJECTIVES

Three-dimensional (3D) cell cultures have many applications such as stem cell biology research, new drug discovery, cancer, and Chronic Rhinosinusitis with Nasal Polyps (CRSwNP). This disease is characterized by a significant impact on quality of life and productivity. The diversity of factors that act in the progression of CRSwNP point to the creation of a cell culture model that allows the integration of different cell types with extracellular matrix. This work aimed to create a cell culture model in 3 dimensions (spheroids) for the study of Nasal Polyposis.

METHODS

Nasal polyp tissue from patients diagnosed with CRSwNP was mechanically dissociated using tweezers and a scalpel and the solution containing cells and small aggregates of nasal polyps was transferred to a Petri dish containing 5 mL of culture medium at the concentration of 10 cells/mL.

RESULTS

The spheroids were cultivated for 20 days, fixed and analyzed using confocal microscopy. In a 3D culture environment, the spheroids were formed both by clustering cells and from small tissue fragments. In the cultures analyzed, the ciliary beat was present from the dissociation of the cells up to 20 days in culture.

CONCLUSION

Our findings also point to these characteristics showing the environment generated in our study, the cells remained differentiated for a longer time and with ciliary beating. Thus, this work shows that nasal polyp-derived cells can be maintained in a 3D environment, enabling better strategies for understanding CRSwNP in situations similar to those found in vivo.

LEVEL OF EVIDENCE

Laboratory studies.

摘要

目的

三维(3D)细胞培养在干细胞生物学研究、新药发现、癌症和慢性鼻-鼻窦炎伴鼻息肉(CRSwNP)等方面有许多应用。这种疾病的特点是对生活质量和生产力有重大影响。作用于 CRSwNP 进展的多种因素表明需要创建一种细胞培养模型,该模型允许不同细胞类型与细胞外基质整合。这项工作旨在创建用于研究鼻息肉的 3 维(球体)细胞培养模型。

方法

使用镊子和手术刀对患有 CRSwNP 的患者的鼻息肉组织进行机械解离,将含有细胞和小鼻息肉组织碎片的溶液转移到含有 5mL 培养基的培养皿中,细胞浓度为 10 个细胞/mL。

结果

将球体培养 20 天,用共聚焦显微镜固定和分析。在 3D 培养环境中,球体的形成既可以通过细胞聚集,也可以通过小块组织片段形成。在分析的培养物中,从细胞解离到培养 20 天,纤毛摆动一直存在。

结论

我们的研究结果还表明,这些特征表明我们研究中产生的环境使细胞保持分化状态更长时间,并保持纤毛摆动。因此,这项工作表明,鼻息肉来源的细胞可以在 3D 环境中维持,从而为在体内类似情况下更好地理解 CRSwNP 提供了更好的策略。

证据水平

实验室研究。

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