Expression data from mouse limb tendon cells during development.

We have undertaken a screen of mouse limb tendon cells in order to identify molecular pathways involved in tendon development. Mouse limb tendon cells were isolated based on Scleraxis (Scx) expression at different stages of development: E11.5, E12.5 and E14.5 Microarray comparisons were carried out between tendon progenitor and differentiated stages. Forelimbs from E11.5, E12.5 and E14.5 Scx-GFP embryos were collected and dissociated with trypsin to obtain cell suspensions. Scx-positive tendon cells were isolated by FACS. RNA was extracted and Fragmented biotin-labelled cRNA samples were hybridized on Affymetrix Gene Chip Mouse Genome 430 2.0 arrays.本研究针对小鼠肢体肌腱细胞开展筛选实验，旨在鉴定参与肌腱发育过程的分子通路。研究人员根据硬化蛋白（Scleraxis, Scx）的表达水平，在胚胎发育的E11.5、E12.5及E14.5三个阶段分离小鼠肢体肌腱细胞，并针对肌腱祖细胞与分化成熟细胞两个阶段开展基因芯片表达谱对比分析。收集上述三个发育阶段的Scx-GFP转基因小鼠胚胎前肢，使用胰蛋白酶解离组织以获得细胞悬液，通过荧光激活细胞分选（Fluorescence Activated Cell Sorting, FACS）分离得到Scx阳性肌腱细胞；随后提取细胞总RNA，制备片段化并经生物素标记的互补RNA（cRNA）样本，将其与Affymetrix Gene Chip Mouse Genome 430 2.0基因芯片进行杂交。