生长方案:	-
处理方案:	stimulation: none; stimulation: CD3/CD28 T Cell Activator (STEMCELL Technologies)
提取方案:	Tissues acquired from donors were maintained in cold saline during transport to the laboratory, typically within 2-4 hours of procurement. We flushed the left lobe of the lungs with cold complete medium (RPMI 1640, 10% FBS, 100 U/ml penicillin, 100 ug/ml streptomycin, 2 mM L-glutamine) and isolated lymph nodes from the hilum, near the intersections of major bronchi and pulmonary veins and arteries, removing all fat. To obtain mononuclear cell suspensions, hilar lymph nodes and the left lateral basal segment of the lung were mechanically processed using a gentleMACS tissue dissociator (Miltenyi Biotec), enzymatically digested (complete medium with 1 mg/ml collagenase D, 1 mg/ml trypsin inhibitor and 0.1 mg/ml DNase for 1 hour at 37℃ in a mechanical shaker) and centrifuged on a density gradient using 30% Percoll Plus (GE Healthcare). We aspirated bone marrow from near the anterior superior iliac crest. For bone marrow and peripheral blood, we isolated mononuclear cells by density gradient centrifugation using Lymphocyte Separation Medium (Corning). Next, we enriched single cell suspensions from all tissues and blood for untouched CD3+ T cells using magnetic negative selection (MojoSort Human CD3+ T cell Isolation Kit; BioLegend). To eliminate any dead cells prior to stimulation, we used a dead cell removal kit (Miltenyi Biotec). We cultured 0.5 -1 x 106 CD3+ enriched cells from each donor tissue for 16 hours at 37℃ in complete medium, with or without TCR stimulation using Human CD3/CD28 T Cell Activator (STEMCELL Technologies). After stimulation, we removed dead cells as above before preparing cells for single-cell RNA-seq.
建库方案:	We loaded single-cell suspensions into a Chromium Single Cell Chip (10x Genomics) according to the manufacturer's instructions for co-encapsulation with barcoded Gel Beads at a target capture rate of ~5,000 individual cells per sample. We barcoded the captured mRNA during cDNA synthesis and converted the barcoded cDNA into pooled single-cell RNA-seq libraries for Illumina sequencing using the Chromium Single Cell 3' Solution (10x Genomics) according to the manufacturer's instructions. We processed all of the samples for a given donor simultaneously with the Chromium Controller (10x Genomics) and prepared the resulting libraries in parallel in a single batch. We pooled all of the libraries for a given donor, each of which was barcoded with a unique Illumina sample index, for sequencing in a single Illumina flow cell.

分子类型:	poly(A)+ RNA
库的片段类型:	PAIRED
库的链类型:	Forward
测序平台:	ILLUMINA
测序仪型号:	Illumina HiSeq 4000
链特异性:	Specific

数据来源	GEN样本编号	GEN数据集编号	系列编号	项目编号	样本编号	样本名称	生物样本编号	样本访问号	实验访问号	释放时间	提交时间	最后更新时间	物种	种族	族裔	年龄	年龄单位	性别	来源名称	组织	细胞类型	细胞亚型	细胞系	疾病	疾病状态	发育阶段	突变/变异	表型	Condition Detail	生长方案	处理方案	提取方案	建库方案	分子类型	库的片段类型	链特异性	库的链类型	加标（Spike-In）	测序方法	测序平台	测序仪型号	细胞数	测序片段数	碱基数	平均测序片段长度_1	平均测序片段长度_2	唯一比对率	多重比对率	覆盖度