Transcriptome analysis of freshly isolated young and aged mouse natural regulatory T cells

Remyelination efficiency declines wiith aged leading to axonal loss and irreversible disability. Regulatory T cells have been previously shown Overall design: Young and aged natural Tregs were isolated from spleen and lymphnodes of C57Bl6/J mice. In brief, spleen and lymph nodes were mashed through a 70µm strainer and red blood cells lysed. Natural Tregs were then magnetically isolated using CD4 negative selection (Stemcell technologies) followed by CD25 positive selection (Stemcell technologies). CD25+ cells were then lysed with Trizol and frozen at -80C. young and aged natural Tregs were always isolated simultaneosly and once all samples were obtained, RNA was isolated.髓鞘再生效率随衰老进程下降，进而导致轴突丢失与不可逆神经功能残疾。已有研究表明调节性T细胞（Regulatory T cells）。实验整体设计如下：从C57Bl6/J小鼠的脾脏与淋巴结中分离年轻及衰老型天然调节性T细胞（natural Tregs）。简言之，将脾脏与淋巴结组织通过70μm细胞筛研磨，并裂解红细胞。随后采用磁珠分选法分离天然调节性T细胞：先通过CD4阴性分选（Stemcell Technologies），再进行CD25阳性分选（Stemcell Technologies）。将分选得到的CD25阳性细胞用Trizol试剂裂解后，于-80℃冻存。年轻与衰老型天然调节性T细胞始终同步分离；待所有样本收集完成后，再进行RNA提取。