Genome-wide ATAC-seq maps in human CD4+ T cells stimulated or unstimulated in vitro. Genome-wide ATAC-seq maps in human CD4+ T cells stimulated or unstimulated in vitro

We report the application of ATAC-seq to CD4+ T cells stimulated with anti-CD3/CD28 T activator beads for four hours in culture versus CD4+ T cells cultured in medium alone. Overall design: CD4+ T cells were enriched from whole blood using the RosetteSep human CD4+ T cell enrichment kit. Enriched cells were stimulated with anto-CD3/CD28 beads at a ratio of 0.3 beads / cell for four hours in X-VIVO + 1% human AB serum and penicillin/streptomycin antibiotic. Non-activated CD4+ T cells were cultured in medium alone for four hours. Nuclei from 50,000 CD4+ T cells were prepared and ATAC-seq libraries generated fo sequencing. Blood was obtained from two unrelated healthy donors recruited through the Cambridge BioResource.

本研究报道了ATAC测序（ATAC-seq）在两组CD4+ T细胞中的应用：一组为采用抗CD3/CD28 T细胞激活磁珠刺激培养4小时的CD4+ T细胞，另一组为仅用基础培养基培养的CD4+ T细胞。整体实验设计如下：研究人员通过RosetteSep人CD4+ T细胞富集试剂盒从全血中分离富集CD4+ T细胞；将富集得到的细胞以0.3磁珠/细胞的比例与抗CD3/CD28磁珠共孵育，在添加1%人AB血清及青霉素/链霉素抗生素的X-VIVO培养基中刺激培养4小时；未激活的CD4+ T细胞则仅使用该培养基培养4小时。从50000个CD4+ T细胞中提取细胞核，并构建ATAC测序（ATAC-seq）文库用于后续测序。本研究的血液样本来源于通过剑桥生物资源库（Cambridge BioResource）招募的两名无亲缘关系的健康志愿者。