Project description:Testis is the most important male reproductive organ, and the integrity of its physiological function is crucial to the successful production of sperm. In this study, the expression profiles of 11 991 and 8 930 cells in testicular tissue of yak and cattle-yak after sexual maturity were established using Single-cell RNA sequencing. The identification results of cell subpopulations and marker genes were analyzed and their possible mechanisms were predicted.

2023-05-22 | GSE225618 | GEO

DMSO cryopreservation: the method of choice for droplet-based single-cell RNA sequencing of preserved cells [10X Genomics]

Project description:Recently, combining single-cell RNA sequencing (scRNA-seq) with upstream cell preservation procedures such as cryopreservation or methanol fixation has become more common. By separating cell handling and preparation, from downstream library generation, scRNA-seq workflows are more flexible and manageable. However, the inherent transcriptomics changes associated with cell preservation and how they may bias further downstream analysis remain unknown. Here, we present a side-by-side droplet-based scRNA-seq analysis, comparing the gold standard – fresh cells – to three different cell preservation workflows: dimethyl sulfoxide based cryopreservation, methanol fixation and CellCover reagent. Cryopreservation proved to be the most robust protocol, maximizing both cell integrity and low background ambient RNA. Importantly, gene expression profiles from fresh cells correlated most with those of cryopreserved cells. Such similarities were consistently observed across the tested cell lines (R ≥ 0.97), monocyte-derived macrophages (R = 0.97) and immune cells (R = 0.99). In contrast, both methanol fixation and CellCover preservation showed an increased ambient RNA background and an overall lower gene expression correlation to fresh cells. Thus, our results demonstrate the superiority of cryopreservation over other cell preservation methods. We expect our comparative study to provide single cell omics researchers invaluable support when integrating cell preservation into their scRNA-seq studies.

2019-07-25 | GSE127248 | GEO

10X genomics single cell sequencing of sheep testis

Project description:Sheep testes undergo a dramatic rate of development with structural changes during sexual maturity, including the proliferation and maturation of somatic niche cells and the initiation of spermatogenesis. To explore this complex process, 12,843 testicular cells from three sexual maturity (3 month-old) rams were sequenced using 10x Genomic platform single-cell sequencing (scRNA-seq).Nine testicular somatic cell types and five male germ cell types were observed.The study revealed significant changes in germline stem cells during sexual maturation. Candidate factors and pathways for the regulation of germ and somatic cell development were identified that represent the scientific basis for the development of a livestock stem cell breeding program.

2021-12-30 | GSE184343 | GEO

10X Genomics - Evaluation of single-Cell RNA Sequencing Platforms

Project description:comparison of commercially available scRNA-Seq instrumentation using a well-defined cell line and treatment for evaluation

2021-10-06 | GSE142647 | GEO

Single-cell RNA-sequencing of bovine fetal gonads using 10x Genomics

Project description:The germ cell lineage ensures the creation of new individuals perpetuating the genetic information across generations. Primordial germ cells (PGCs) are pioneers of gametes and exist transiently during development until they differentiate into oogonia in females, or spermatogonia in males. Little is known about the molecular characteristics of PGCs in cattle. By performing single-cell RNA-sequencing of bovine fetal gonads we evaluated the molecular signatures of bovine germ cells and their niche at 50 days of development. Our results indicate that PGCs and somatic cells were in very early stages of development, and that bovine and human germ cells share expression of transcriptional regulators of germline specification, and several genes coding for membrane-bound proteins. Additionally, the expression of members of Notch, Nodal/Activin, and BMP signaling cascades in the bovine fetal ovary, suggests that these pathways are involved in the interaction between germ cells and their niche. Results of this study provide insights into the mechanisms involved in the development of bovine PGCs and put in evidence similarities between the bovine and human germline.

2021-01-29 | GSE162952 | GEO

Single cell RNA-sequencing of Spike-ins and Control RNA using 10x Genomics Chromium system

Project description:In this study, we assess technical differences between commonly used single-cell RNA-Sequencing (scRNA-Seq) methods. In this dataset, we assess the RNA detection rates using high-throughput 10x Genomics Chromium system. We mix equal volume of Control Brain RNA (3ul; FirstChoice Total Brain RNA; #AM7962) and ERCC spikes (3ul 1:4 dilution; #4456653) to make a '2x Control RNA+ERCC' master mix. The '2x Control RNA+ERCC' master mix is diluted with equal volume nuclease-free water to make '1x Control RNA+ERCC' master mix. 3ul of '1x Control RNA+ERCC' master mix is added to Chromium single cell suspension and processed as per Chromium guidelines. The sample-data relationship format (SDRF) file for this submission contains only a high-level representation of the sample, library and run information per flow cell, and not per cell. For meta-data at the level of individual cells, please refer to the supplementary file called single_cells_list.txt, which is included as part of this ArrayExpress submission.

2017-03-08 | E-MTAB-5480 | biostudies-arrayexpress

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