Choosing the right chondrocyte cell line: Focus on nitric oxide
Choosing the right chondrocyte cell line: Focus on nitric oxide
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DOI：
10.1002/jor.22954
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发表时间：
2015-12-01
影响因子：
2.8
通讯作者：
Gualillo, Oreste
中科院分区：
文献类型：
--
作者：
Santoro, Anna;Conde, Javier;Gualillo, Oreste
一氧化氮(NO)被认为是一种分解代谢因子，通过诱导软骨细胞的凋亡、基质金属蛋白酶的合成和促炎细胞因子的表达而参与了骨关节炎的病理过程。因此，软骨细胞中NO调控的研究是一个需要深入探讨的相关领域。然而，到目前为止，只有小鼠ATDC-5细胞系和原代软骨细胞是研究软骨组织中NO产生的成熟细胞。本研究的目的是确定两个常用的人软骨细胞系：SW-1353和T/C-28a2细胞系是否是检测内毒素和/或促炎细胞因子驱动的NO释放和iNOS表达的良好模型。为此，我们仔细研究了内毒素和白介素1单独或联合刺激的人T/C-28a2和SW-1353软骨细胞的NO产生和iNOS蛋白表达。我们还使用ATDC-5细胞作为NO产生的阳性对照。通过比色法Griess反应确定NO的积累，而通过Western Blot分析确定NOS II型的表达。我们的结果清楚地表明，无论是T/C-28a2还是SW-1353软骨细胞，在细菌内毒素或细胞因子用IL-1攻击后，都没有检测到NO产生或iNOS表达的增加。我们的研究表明T/C-28a2和SW-1353细胞不适合研究NO的释放和iNOS的表达，证实了ATDC5和人原代培养的软骨细胞是研究该介质的作用的最佳体外细胞系统。(C)2015年骨科研究会。由Wiley期刊公司出版，J Orthop Res 33：1784-1788,2015。
Nitric oxide (NO) has been considered a catabolic factor that contributes to OA pathology by inducing chondrocytes apoptosis, matrix metalloproteinases synthesis, and pro-inflammatory cytokines expression. Thus, the research on NO regulation in chondrocytes represents a relevant field which needs to be explored in depth. However, to date, only the murine ATDC-5 cell line and primary chondrocytes are well-established cells to study NO production in cartilage tissues. The goal of this study is to determine whether two commonly used human chondrocytic cell lines: SW-1353 and T/C-28a2 cell lines are good models to examine lipopolysaccharide and/or pro-inflammatory cytokine-driven NO release and iNOS expression. To this aim, we carefully examined NO production and iNOS protein expression in human T/C-28a2 and SW-1353 chondrocytes stimulated with LPS and interleukin (IL)-1 alone or in combination. We also use ATDC-5 cells as a positive control for NO production. NO accumulation has been determined by colorimetric Griess reaction, whereas NOS type II expression was determined by Western Blot analysis. Our results clearly demonstrated that neither human T/C-28a2 nor SW-1353 chondrocytes showed a detectable increase in NO production or iNOS expression after bacterial endotoxin or cytokines challenge with IL-1. Our study demonstrated that T/C-28a2 and SW-1353 human cell lines are not suitable for studying NO release and iNOS expression confirming that ATDC5 and human primary cultured chondrocytes are the best in vitro cell system to study the actions derived from this mediator. (c) 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:1784-1788, 2015.