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早幼粒细胞系中CD14表达的诱导及向单核细胞或成熟巨噬细胞的分化:新方法

Induction of CD14 Expression and Differentiation to Monocytes or Mature Macrophages in Promyelocytic Cell Lines: New Approach.

作者信息

Zamani Fatemeh, Zare Shahneh Fateme, Aghebati-Maleki Leili, Baradaran Behzad

机构信息

Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. ; Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Adv Pharm Bull. 2013;3(2):329-32. doi: 10.5681/apb.2013.053. Epub 2013 Aug 20.

DOI:10.5681/apb.2013.053
PMID:24312856
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3848216/
Abstract

PURPOSE

CD14, one of the main differentiation markers on the surface of myeloid lineage cells, acts as a key role in activation of LPS-induced monocytes. LPS (lipopolysaccharide) binds to LPS-binding protein in plasma and are delivered to the cell surface receptor CD14. In this study, Various stimuli [Dimethyl Sulfoxide (DMSO), active 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and LPS], either alone or in combination, have been recognized that have an effect on the level of CD14 expression in the human HL-60 and U937 promonocytic cell lines and therefore induce their terminal differentiation into monocytes or mature macrophages.

METHODS

U937 and HL-60 cells were cultured in RPMI 1640 supplemented with 10% FBS. For each cell line, 1Ã106 cells were seeded for 72 hours with DMSO, 14 days with LPS and 18 days with 1, 25-D3 in each well plate; then ELISA method was used to study their responses to the factors by means of anti-CD14.

RESULTS

ELISA assay demonstrated that U937 and HL-60 cells were induced by both [1,25(OH)2D3] and DMSO to obtain characteristics of adherent cells and express CD14 protein; moreover, LPS at a low dose increased CD14 expression on surface of this cells.

CONCLUSION

According to the our results, it is speculated that CD14 gene expression may be induced in human U937 and HL-60 cell lines by different factors including 1,25-D3, DMSO and LPS.

摘要

目的

CD14是髓系细胞表面主要的分化标志物之一,在脂多糖(LPS)诱导的单核细胞激活中起关键作用。LPS与血浆中的LPS结合蛋白结合,并被递送至细胞表面受体CD14。在本研究中,已认识到各种刺激因素[二甲基亚砜(DMSO)、活性1,25 - 二羟基维生素D3 [1,25(OH)2D3]和LPS]单独或联合使用时,会对人HL - 60和U937原单核细胞系中CD14的表达水平产生影响,从而诱导它们终末分化为单核细胞或成熟巨噬细胞。

方法

U937和HL - 60细胞在补充有10%胎牛血清的RPMI 1640培养基中培养。对于每个细胞系,在每个孔板中接种1×10⁶个细胞,分别用DMSO培养72小时,用LPS培养14天,用1,25 - D3培养18天;然后采用ELISA法,通过抗CD14来研究它们对这些因素的反应。

结果

ELISA分析表明,[1,25(OH)2D3]和DMSO均可诱导U937和HL - 60细胞获得贴壁细胞特性并表达CD14蛋白;此外,低剂量的LPS可增加这些细胞表面CD14的表达。

结论

根据我们的结果推测,包括1,25 - D3、DMSO和LPS在内的不同因素可能诱导人U937和HL - 60细胞系中CD14基因的表达。

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