An Organ Culture System to Model Early Degenerative Changes of the Intervertebral Disc: II Profiling Global Gene Expression Changes. Rattus norvegicus

To mimic a degenerative insult, rat intervertebral discs were cultured in the presence of TNF-alpha, IL-1beta and serum limiting conditions. Gene expression analysis was performed using a microarray to identify differential gene expression between experimental and control groups. Overall design: The discs were collected and the annulus fibrosus (AF) was separated from the nucleus pulposus (NP). Total RNA from NP was used for RNA extraction and hybridization on Affymetrix microarrays: Control group - Ctr-1, Ctr-2, Ctr-3, Ctr-4 and Experimental group-Exp-1, Exp-2, Exp-3, Exp-4.

为模拟退行性损伤，我们在肿瘤坏死因子-α（TNF-alpha）、白细胞介素-1β（IL-1beta）及血清限制培养条件下对大鼠椎间盘进行体外培养。本研究采用基因芯片（microarray）开展基因表达分析，以鉴定实验组与对照组间的差异基因表达谱。
整体实验设计：收集大鼠椎间盘后，将纤维环（annulus fibrosus, AF）与髓核（nucleus pulposus, NP）分离；从髓核中提取总RNA，用于Affymetrix基因芯片的杂交检测。对照组包含Ctr-1、Ctr-2、Ctr-3、Ctr-4四个样本，实验组包含Exp-1、Exp-2、Exp-3、Exp-4四个样本。