Project description:To reveal distinct transcriptomes associated with spermatogonial stem cell renewal vs. initiation of differentiation, single-cell transcriptomes from P6 ID4-EGFP+ spermatogonia were subdivided into subpopulations that displayed distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the Fluidigm C1 instrument to capture individual spermatogonia for SMART-Seq2 single-cell RNA-seq.
Project description:To reveal distinct transcriptomes associated with spermatogonial stem cell renewal vs. initiation of differentiation, single-cell transcriptomes from P6 ID4-EGFP+ spermatogonia (sorted for brightest or dimmest) or unselected testis cells were used for Drop-Seq analysis. The GFP-bright and dim phenotypes exhibit distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the 10x Genomics Chromium (a commercial Drop-Seq variant) to perform single-cell RNA-seq
Project description:To reveal distinct transcriptomes associated with spermatogonial stem cell renewal vs. initiation of differentiation, single-cell transcriptomes from Adult ID4-EGFP+ spermatogonia were subdivided into subpopulations that displayed distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the Fluidigm C1 instrument to capture individual spermatogonia for SMART-Seq2 single-cell RNA-seq.
Project description:P6 ID4-EGFP+ undifferentiated spermatogonia, including those stained robustly (high) or weakly (low) for TSPAN8 were isolated by FACS.
Project description:Purpose: We performed a time-course single-cell RNA-seq of the somatic cells of the XX mouse gonads to study the cell population heterogeneity and the genetic program during their differentiation. Methods: We collected gonads from NR5A1-eGFP transgenic embryos at six embryonic stages: E10.5, E11.5, E12.5, E13.5, E16.5 and P6. Methods: Cells were capture with the C1 autoprep system and cDNA sequenced with Illumina HiSeq 2000. Results: One cell population was detected at E10.5 and give rise to both Granulosa and steroidogenic precursor cells. A precursor cell population remains undifferentiated at P6 and are likely to be theca cell precursors. Conclusion: Our study is, to date, the most granular transcriptomic study of the developing mouse ovary and provide a more complete model of somatic cell differentiation during female sex determination.
Project description:To reveal distinct transcriptome changes among ID4-EGFP-bright adult mouse spermatogonia associated with mTORC1 activity, single-cell transcriptomes were generated from GFP-bright/CD9-bright spermatogonia from adult mice in three groups: control (untreated), 2 days of Rapamycin treatment (Rapamycin) and 2 days Rapamycin plus 1 day washout (Rapamycin_Release). Based on transplantation studies performed previously, ID4-EGFPbright cells are highly enriched for SSCs. We used the 10x Genomics Chromium to perform single-cell RNA-seq.
Project description:To reveal distinct transcriptomes associated with various spermatogenic cells, including spermatogonial stem cells and all of their subsequent progeny, single-cell transcriptomes from Adult ID4-EGFP+ spermatogonia (sorted for brightest or dimmest), StaPut-enriched spermatocytes and spermatids, or unselected steady-state spermatogenic cells were used for 10x Genomics analysis. The GFP-bright and dim phenotypes exhibit distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the 10x Genomics Chromium (Drop-Seq) to perform single-cell RNA-seq