PRJNA554092: The Dynamic Transcriptional Cell Atlas of Testis Development During Human Puberty
概要: The human testis undergoes dramatic developmental and structural changes during puberty, including proliferation and maturation of niche/somatic cells, and the onset of spermatogenesis. Here, we profiled and analyzed single-cell transcriptomes of ~10,000 testicular cells from four boys spanning puberty, and compared to infants and adults. During puberty, undifferentiated spermatogonia first expand and then differentiate, prior to gametogenesis. Notably, we identify a common pre-pubertal progenitor for Leydig and myoid cells, and reveal candidate factors/pathways for pubertal differentiation. Furthermore, pre-pubertal Sertoli cells form two states that differ in mitochondrial/metabolic transcription, which converge to a single mature population during puberty. Roles for testosterone in Sertoli cell maturation, antimicrobial peptide secretion and spermatogonial differentiation are revealed through analyses of testosterone-suppressed transgender female testis and via in vitro seminiferous tubule culturing. Overall, our transcriptional atlas of the developing human testis provides major insights into developmental changes and key factors/pathways that accompany male puberty.
项目整体设计: Single-cell RNA-seq of testicular cells from different ages of boys in puberty. Testosterone-suppressed transgender female testis samples are included.
| 生长方案: |
- |
| 处理方案: |
- |
| 提取方案: |
Testicular tissues were firstly minced into small pieces mechanically, and subject to trypsin digestion for ~25 min at 37 C. Cell suspension was centrifuged and resuspended in cold PBS buffer with 0.4% of BSA at the concentration of ~1000/ul, which then was loaded onto the 10X Chromium Controller using Chromium Single Cell 3' v2 reagents. |
| 建库方案: |
Sequencing library was prepared following the manufacturer's instructions (10X Genomics), with 13 cycles used for cDNA amplification and 12 cycles for library amplification, which was then sequenced on a 26 X 100 cycle paired-end run via a Illumina 2500 instrument. Each sample was sequenced in one separate sequencing lane. |
| 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 2500 |
| 链特异性: |
Specific |
| 数据来源 |
GEN样本编号 |
GEN数据集编号 |
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样本编号 |
样本名称 |
生物样本编号 |
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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 |
The Dynamic Transcriptional Cell Atlas of Testis Development during Human Puberty.
Cell stem cell . 2020-01-09 [PMID:
31928944]