Dissect formative pluripotency using cultured pluripotent stem cells

使用培养的多能干细胞剖析形成性多能性

• 批准号: 10621933
• 负责人: Jun Wu
• 金额: $ 34.06万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-22 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621933
• 关键词: Adult; Affinity Chromatography; Cell Differentiation process; Cell Lineage; Cells; ChIP-seq; Characteristics; Chemicals; Chimera organism; Clustered Regularly Interspaced Short Palindromic Repeats; Competence; Derivation procedure; Development; Developmental Biology; Embryo; Epiblast; Fibroblast Growth Factor; Fibroblast Growth Factor Receptors; Foundations; Gastrula; Gene Expression Profile; Generations; Genetic; Germ Cells; Goals; Homeodomain Proteins; Human; In Situ; In Vitro; Infertility; Laboratories; Maintenance; Methods; Modeling; Molecular; Mus; Names; Oocytes; Oogonia; Pathway interactions; Phase; Phenotype; Play; Pluripotent Stem Cells; Proteins; Proteomics; Regenerative Medicine; Regulation; Regulatory Element; Reproductive Biology; Resources; Role; Signal Transduction; Somatic Cell; Specific qualifier value; Structure of primordial sex cell; Study models; Testing; Time; Tissues; Transforming Growth Factor beta; Transgenes; Validation; austin; blastocyst; cell type; embryonic stem cell; established cell line; experimental study; implantation; in vivo; induced pluripotent stem cell; insight; novel; permissiveness; pluripotency; regeneration potential; stem cell model; stem cells; tool; transcription factor; transcriptome

PROJECT SUMMARY/ABSTRACT Derivation of pluripotent stem cells (PSCs) has revolutionized developmental biology and regenerative medicine. To stably maintain PSCs in culture and guide them to differentiate with high efficiency and fidelity into a variety of cell types, it is important to understand the molecular mechanisms governing pluripotency (the ability of a cell to generate any tissues in the body). Two phases of pluripotency, naïve and primed, have been defined and studied in detail thanks to the successful derivation of mouse embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs), respectively. Mouse ESCs most closely resemble epiblast from a 4-day-old mouse blastocyst (~embryonic day 4, or E4), while “primed” EpiSCs display a global gene expression signature similar to the E7 epiblast of a post-implantation mouse embryo. Despite these advances, however, however, there is lack of a well-established PSC model that resembles E5-6 early post-implantation epiblast, which corresponds to the formative phase of pluripotency. Formative pluripotency exists within a time window during which naïve pluripotency is reconfigured to prepare for multilineage competency, including germ cells. Functionally, formative pluripotency is characterized by both chimera competency and permissiveness for direct primordial germ cell (PGC) induction. Several recent studies have attempted to define this state by transient epiblast-like cells (EpiLCs) differentiated from ESCs. To date, however, stable formative PSCs have not yet been generated. By modulating the FGF, TGF-β and WNT pathways, we recently derived PSCs from both mice and humans (referred to as FTW-PSCs) that are permissive for direct PGC-like cell induction in vitro and are capable of contributing to intra- or inter-species chimeras in vivo. FTW-PSCs harbor molecular, cellular and phenotypic features characteristic of formative pluripotency. The overall objective of this proposal is to use these newly established cell lines to comprehensively dissect the formative state across species. The proposed studies will elucidate the roles of several transcription factors in regulating mouse and human formative pluripotency, as well as demonstrate that FTW-PSCs are a robust platform for dissecting the molecular mechanisms underlying human and mouse PGC specification. In addition, we will establish an in vitro platform for the generation of functional mouse oocytes and human oogonia based on formative FTW-PSCs, thereby providing an invaluable resource for studying germ cell development and human infertility. Our proposal has tremendous potential to revolutionize regenerative medicine and reproductive biology.

项目总结/文摘

项目成果

Self-organization of vascularized skeletal muscle from bovine embryonic stem cells.

来自牛胚胎干细胞的血管化骨骼肌的自组织。

• DOI: 10.1101/2024.03.22.586252
• 发表时间: 2024
• 期刊: bioRxiv : the preprint server for biology
• 作者: Sanaki-Matsumiya,Marina;Villava,Casandra;Rappez,Luca;Haase,Kristina;Wu,Jun;Ebisuya,Miki
• 通讯作者: Ebisuya,Miki