Pluripotent stem cells related to embryonic disc exhibit common self-renewal requirements in diverse livestock species
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PROVIDER: PRJNA723284 | ENA |
REPOSITORIES: ENA
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Project description:Despite four decades of effort, robust propagation of pluripotent stem cells from livestock animals remains challenging. The requirements for self-renewal are unclear and the relationship of cultured stem cells to pluripotent cells resident in the embryo uncertain. Here we avoided feeder cells or serum factors to provide a defined culture microenvironment. We show that the combination of Activin A, Fibroblast growth factor, and Wnt inhibitor XAV939 (AFX), supports establishment and continuous expansion of pluripotent stem cell lines from porcine, ovine and bovine embryos. Germ layer differentiation was evident in teratomas and readily induced in vitro. Global transcriptome analyses highlighted commonality in transcription factor expression across the three species, while global comparison with porcine embryo stages showed proximity to bilaminar disc epiblast. Clonal genetic manipulation and gene targeting were exemplified in porcine stem cells. We further demonstrated that genetically modified AFX stem cells gave rise to cloned porcine foetuses by nuclear transfer. In summary, for major livestock mammals pluripotent stem cells related to the formative embryonic disc are reliably established using a common and defined signaling environment.
2021-11-15 | GSE172420 | GEO
Project description:Despite four decades of effort, robust propagation of pluripotent stem cells from livestock animals remains challenging. The requirements for self-renewal are unclear and the relationship of cultured stem cells to pluripotent cells resident in the embryo uncertain. Here, we avoided using feeder cells or serum factors to provide a defined culture microenvironment. We show that the combination of activin A, fibroblast growth factor and the Wnt inhibitor XAV939 (AFX) supports establishment and continuous expansion of pluripotent stem cell lines from porcine, ovine and bovine embryos. Germ layer differentiation was evident in teratomas and readily induced in vitro. Global transcriptome analyses highlighted commonality in transcription factor expression across the three species, while global comparison with porcine embryo stages showed proximity to bilaminar disc epiblast. Clonal genetic manipulation and gene targeting were exemplified in porcine stem cells. We further demonstrated that genetically modified AFX stem cells gave rise to cloned porcine foetuses by nuclear transfer. In summary, for major livestock mammals, pluripotent stem cells related to the formative embryonic disc are reliably established using a common and defined signalling environment. This article has an associated 'The people behind the papers' interview.
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Project description:Nosala C, Hagen KD, Hilton N, et al. Disc-associated proteins (DAPs) mediate the unusual hyperstability of Giardia's ventral disc. J Cell Sci. 2020; ejcs.227355.
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Project description:The biological characteristics of the temporomandibular joint (TMJ) disc involve complex changes in cell identity and extracellular matrix (ECM) composition to modulate jaw function during postnatal growth and aging. To better understand this process, we profiled and analyzed 39111 single-cell transcriptomes of TMJ disc cells from mice at different postnatal stages. Our data showed that TMJ disc cells are composed of 9 distinct clusters that can be divided into 4 principal cell types: fibroblasts (FBs), which were the most abundant, endothelial cells (ECs), macrophages (MPhs) and mural cells (MCs). FBs were found to exhibit heterogeneity in terms of biological processes related to ECM secretion and metabolic functions. The ECs and MPhs mainly clustered together in the junction of the anterior disc band and anterior attachment, indicating the potential importance of the inflammatory response in this region in the pathological process of disc diseases. Notably, we found for the first time that resident MCs are the source of the progenitor niche in TMJ discs. The pseudotime trajectory showed that MCs have 2 lineage fates: self-renewal or multidirectional differentiation toward functional cells. In MCs, the NOTCH3 and THY1 signaling pathways were ubiquitously active at all stages, coupled with specific gene expression of Notch3 and Thy1 in MCs, indicating the critical role of these molecules in MC characteristics. As an example, THY1+ cells in TMJ discs showed markedly stronger stem cell characteristics than THY1- disc cells in vitro and ex vivo. In addition, lineage tracing in vivo showed that the Myh11-CreER+ MC lineage proliferated and migrated to coordinate angiogenesis during disc injury but gradually lost its progenitor characteristics and ultimately became a functional cell for disc repair. Overall, we reveal multiple insights into the coordinated and temporal development of TMJ disc cells at different postnatal stages and are the first to describe the resident MC progenitor cell niche during TMJ disc growth and injury.
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Project description:DNA methylation is an important epigenetic modification that is widely conserved across animal genomes. It is widely accepted that DNA methylation patterns can change in a context-dependent manner, including in response to changing environmental parameters. However, this phenomenon has not been analyzed in animal livestock yet, where it holds major potential for biomarker development. Building on the previous identification of population-specific DNA methylation in clonal marbled crayfish, we have now generated numerous base-resolution methylomes to analyze location-specific DNA methylation patterns. We also describe the time-dependent conversion of epigenetic signatures upon transfer from one environment to another. We further demonstrate production system-specific methylation signatures in shrimp, river-specific signatures in salmon and farm-specific signatures in chicken. Together, our findings provide a detailed resource for epigenetic variation in animal livestock and suggest the possibility for origin tracing of animal products by epigenetic fingerprinting.
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