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传代和形态对颞下颌关节盘细胞基因表达的影响。

Effect of passage and topography on gene expression of temporomandibular joint disc cells.

作者信息

Allen Kyle D, Athanasiou Kyriacos A

机构信息

Department of Bioengineering, Rice University, Houston, Texas 77251-1892, USA.

出版信息

Tissue Eng. 2007 Jan;13(1):101-10. doi: 10.1089/ten.2006.0094.

Abstract

The temporomandibular joint (TMJ) disc is maintained by a population of fibrochondrocytes. Although articular chondrocytes exhibit zonal differences and de-differentiate in monolayer culture, such variations are unknown for fibrochondrocytic populations. This study's objective was to define topographical cellular variations in the porcine TMJ disc and investigate changes in the disc's gene expression levels over multiple passages using quantitative reverse transcriptase polymerase chain reaction. For topographical characterization, samples were acquired from posterior, anterior, lateral, medial, and intermediate zone sections and subdivided into inferior and superior halves. For passage characterization, cells were plated and passaged for 35 days, with samples acquired at every passage. The medial region had the lowest expression of genes indicative of fibroblastic activity, but in general, topographical variations were limited. Passage effects were evident; gene expression levels of aggrecan, collagen type I, and collagen type II dropped 20%, 23%, and 73% per passage, respectively. In contrast, decorin and glyseraldehyde-3-phosphate dehydrogenase gene expression increased 33% and 27% per passage, respectively. These data indicate that TMJ disc cells undergo significant changes due to monolayer expansion, experiencing losses in major chondrocytic markers (aggrecan and collagen type II) and fibroblastic markers (collagen type I) and posing a serious impediment to studies in which cell passaging is required.

摘要

颞下颌关节(TMJ)盘由一群纤维软骨细胞维持。尽管关节软骨细胞在单层培养中表现出区域差异并去分化,但对于纤维软骨细胞群体而言,这种变化尚不清楚。本研究的目的是确定猪TMJ盘的地形学细胞变化,并使用定量逆转录聚合酶链反应研究多代培养过程中盘的基因表达水平变化。为了进行地形学特征分析,从后、前、外侧、内侧和中间区域切片获取样本,并将其细分为下半部和上半部。为了进行传代表征,将细胞接种并传代培养35天,每一代都获取样本。内侧区域中指示成纤维细胞活性的基因表达最低,但总体而言,地形学变化有限。传代效应明显;每传一代,聚集蛋白聚糖、I型胶原蛋白和II型胶原蛋白的基因表达水平分别下降20%、23%和73%。相比之下,核心蛋白聚糖和甘油醛-3-磷酸脱氢酶基因表达每传一代分别增加33%和27%。这些数据表明,TMJ盘细胞因单层扩增而发生显著变化,主要软骨细胞标志物(聚集蛋白聚糖和II型胶原蛋白)和成纤维细胞标志物(I型胶原蛋白)减少,这对需要细胞传代的研究构成严重障碍。

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