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基于外周血血浆来源纤维蛋白凝胶建立U266细胞系的三维培养条件

Establishment of A Three-Dimensional Culture Condition for The U266 Cell Line Based on Peripheral Blood Plasma-Derived Fibrin Gels.

作者信息

Jomehpour Mahshid, Khosravi Mohammadmahdi, Janfada Mehrnaz, Abroun Saeid, Vahdat Sadaf

机构信息

Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Applied Cell Sciences Division, Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Email:

出版信息

Cell J. 2023 Apr 1;25(4):229-237. doi: 10.22074/cellj.2023.562849.1138.

Abstract

OBJECTIVE

The study of pathophysiology as well as cellular and molecular aspects of diseases, especially cancer, requires appropriate disease models. three-dimensional (3D) structures attracted more attention to recapitulate diseases rather than in vitro two-dimensional (2D) cell culture conditions because they generated more similar physiological and structural properties. Accordingly, in the case of multiple myeloma (MM), the generation of 3D structures has attracted a lot of attention. However, the availability and cost of most of these structures can restrict their use. Therefore, in this study, we aimed to generate an affordable and suitable 3D culture condition for the U266 MM cell line.

MATERIALS AND METHODS

In this experimental study, peripheral blood-derived plasma was used to generate fibrin gels for the culture of U266 cells. Moreover, different factors affecting the formation and stability of gels were evaluated. Furthermore, the proliferation rate and cell distribution of cultured U266 cells in fibrin gels were assessed.

RESULTS

The optimal calcium chloride and tranexamic acid concentrations were 1 mg/ml and 5 mg/ml for gel formation and stability, respectively. Moreover, the usage of frozen plasma samples did not significantly affect gel formation and stability, which makes it possible to generate reproducible and available culture conditions. Furthermore, U266 cells could distribute and proliferate inside the gel.

CONCLUSION

This available and simple fibrin gel-based 3D structure can be used for the culture of U266 MM cells in a condition similar to the disease microenvironment.

摘要

目的

对疾病,尤其是癌症的病理生理学以及细胞和分子层面进行研究,需要合适的疾病模型。三维(3D)结构比体外二维(2D)细胞培养条件更能吸引人们对疾病模拟的关注,因为它们能产生更相似的生理和结构特性。因此,在多发性骨髓瘤(MM)的情况下,3D结构的构建引起了广泛关注。然而,这些结构中的大多数的可用性和成本可能会限制它们的使用。所以,在本研究中,我们旨在为U266骨髓瘤细胞系创建一种经济实惠且合适的3D培养条件。

材料与方法

在本实验研究中,使用外周血来源的血浆生成纤维蛋白凝胶用于培养U266细胞。此外,评估了影响凝胶形成和稳定性的不同因素。此外,还评估了U266细胞在纤维蛋白凝胶中的增殖率和细胞分布。

结果

氯化钙和氨甲环酸的最佳浓度分别为1mg/ml和5mg/ml,用于凝胶形成和稳定性。此外,使用冷冻血浆样本对凝胶形成和稳定性没有显著影响,这使得创建可重复且可用的培养条件成为可能。此外,U266细胞可以在凝胶内分布和增殖。

结论

这种基于纤维蛋白凝胶的可用且简单的3D结构可用于在类似于疾病微环境的条件下培养U266骨髓瘤细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a912/10201360/35b98e37018a/Cell-J-25-229-g01.jpg

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