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用黑色素瘤或激活和非激活的 Jurkat-E6 细胞产生的条件培养基抑制鼠破骨细胞生成。

Murine osteoclastogenesis suppression using conditioned media produced by melanoma or activated and non-activated Jurkat-E6 cells.

机构信息

Laboratory of Genetics, Butantan Institute, São Paulo, Brazil.

Center of Excellence in New Target Discovery (CENTD), Butantan Institute, São Paulo, Brazil.

出版信息

Cell Biol Int. 2020 May;44(5):1184-1192. doi: 10.1002/cbin.11317. Epub 2020 Feb 20.

Abstract

Conditioned medium (CM) (cell secretome) is a cocktail of growth factors, cytokines, and other soluble mediators secreted by cells into a culture medium. These growth factors are fundamental in many cellular processes such as cell growth, differentiation, and others and the composition of these factors is individual for each cell type. Osteoclasts are large multinucleated cells that are responsible for bone resorption. Immune and cancer cells are known to produce different growth factors, which are able to induce or inhibit osteoclast differentiation. Herein, we evaluated the effect of CM obtained from the supernatant of activated and non-activated Jukart-E6 cells, as well as from one murine (B16-F10) and one human melanoma cell line (SK-MEL-28). To induce osteoclast differentiation, murine bone marrow mononuclear cells were cultured in the presence and absence of differentiation factors (DF), such as macrophage colony-stimulating factor, prostaglandin E2, receptor activator of nuclear factor-κB ligand, and CM. We measured the concentration of interleukin 6, tumor necrosis factor-α and interferon γ (IFN-γ) in CM that can inhibit or induce osteoclastogenesis. Our study demonstrated that CM obtained from each cell line suppresses or inhibits osteoclasts formation at early and intermediate stages of differentiation in the absence or presence of DF. CM obtained from activated Jurkat-E6 cells demonstrates a stronger effect when compared with CM from naïve Jurkat-E6 cells or human and murine melanoma cells. Moreover, CM obtained from activated Jurkat-E6 cells shows higher secretion of IFN-γ, which is an inhibitor of osteoclastogenesis, in comparison with CM obtained from the three other cell lines. On the other hand, CM derived from B16-F10 cells showed a smaller inhibitory effect when compared with CM derived from the other cells.

摘要

条件培养基(CM)(细胞外泌体)是细胞分泌到培养基中的生长因子、细胞因子和其他可溶性介质的混合物。这些生长因子在许多细胞过程中起着重要作用,如细胞生长、分化等,并且这些因子的组成对于每种细胞类型都是独特的。破骨细胞是负责骨吸收的大型多核细胞。已知免疫细胞和癌细胞会产生不同的生长因子,这些生长因子能够诱导或抑制破骨细胞分化。在此,我们评估了从激活和未激活的 Jukart-E6 细胞的上清液中获得的 CM,以及从一种鼠(B16-F10)和一种人黑色素瘤细胞系(SK-MEL-28)获得的 CM 的影响。为了诱导破骨细胞分化,在存在和不存在分化因子(DF)(如巨噬细胞集落刺激因子、前列腺素 E2、核因子-κB 配体受体激活剂)的情况下,将鼠骨髓单核细胞在培养基中培养。我们测量了 CM 中可以抑制或诱导破骨细胞生成的白细胞介素 6、肿瘤坏死因子-α和干扰素 γ(IFN-γ)的浓度。我们的研究表明,从每种细胞系获得的 CM 在不存在或存在 DF 的情况下,在破骨细胞分化的早期和中期阶段均抑制或抑制破骨细胞的形成。与幼稚 Jurkat-E6 细胞或人黑色素瘤和鼠黑色素瘤细胞相比,从激活的 Jurkat-E6 细胞获得的 CM 具有更强的作用。此外,与从其他三个细胞系获得的 CM 相比,从激活的 Jurkat-E6 细胞获得的 CM 显示出更高的 IFN-γ分泌,IFN-γ是破骨细胞生成的抑制剂。另一方面,与从其他细胞获得的 CM 相比,源自 B16-F10 细胞的 CM 显示出较小的抑制作用。

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