Unraveling the Role of Asymmetric Division in Lineage Specification of Stem Cells

揭示不对称分裂在干细胞谱系规范中的作用

• 批准号: 9099760
• 负责人: Ehsan Jabbarzadeh
• 金额: $ 7.33万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9099760
• 关键词: Address; Adherent Culture; Area; Biological Phenomena; Cell Culture Techniques; Cell Lineage; Cell Polarity; Cell Shape; Cell division; Cells; Cues; Cytoskeletal Modeling; Daughter; Disease; Disease model; Embryo; Engineering; Engraftment; Environment; Extracellular Matrix; Generations; Germ Layers; Goals; Health; Homing; Human; Investigation; Maps; Mechanics; Medical; Methods; Mitotic; Mitotic spindle; Modeling; Musculoskeletal Diseases; Natural regeneration; Pattern; Physiological; Pluripotent Stem Cells; Printing; Process; Proliferating; Proteins; Regenerative Medicine; Regulation; Research; Role; Shapes; Signal Transduction; Staging; Stem cells; Techniques; Testing; Tissue Engineering; Tissues; Trauma; Vision; Work; cell type; daughter cell; drug testing; epithelial to mesenchymal transition; in vivo; innovation; insight; monolayer; novel; novel strategies; pluripotency; regenerative therapy; self-renewal; stem cell division; stem cell fate specification; tool

 DESCRIPTION (provided by applicant): There is a profound need for regeneration strategies due to trauma and various musculoskeletal diseases. Human pluripotent stem cells (hPSCs) offer a promising tool for regenerative therapies because of their unique capability to self-renew, proliferate nearly indefinitely, and rise to various cell types. In the regeneration process, the temporary extracellular matrix (ECM) provides multiple signals to hPSCs and guides the process of new matrix formation. Fundamental questions on how hPSCs sense and respond to the cues from surrounding environment await further exploration. A main hurdle facing the studies of hPSC-ECM interactions lies in limitation of hPSCs culture techniques which use clumps of cells from dissected colonies. In these culture methods, hPSCs unavoidably pass through the stage of spontaneous differentiation to all three embryonic germ layers before the generation of the desired tissue-specific cell type. The goal of this study is to generate and characterize a monolayer single-cell culture model that allows for the investigation of biological phenomena associated with hPSC lineage specification. We will evaluate the potential of hPSCs cultured in monolayers to undergo epithelial-to-mesenchymal transition (EMT) and differentiate to specific germ layers. Using the single-cell monolayer culture method, we will explore the role of symmetric/asymmetric cell division in regulation of hPSC pluripotency and lineage specification. This will be accomplished by employing microcontact printing techniques to control cell polarity and cytoskeletal organization of at a single-cell level. This project has wo specific aims: (1) determine the potential of hPSCs cultured in a monolayer to undergo EMT, and (2) determine the role of asymmetric division in fate specification of hPSCs. The novel approach developed in this study has broad applicability to many research areas including tissue engineering and regenerative medicine, disease modeling and drug testing.

 描述（由申请人提供）：由于创伤和各种肌肉骨骼疾病，对再生策略的迫切需要人类多能干细胞（hPSC）由于其独特的自我更新能力，为再生治疗提供了一种有前景的工具。 几乎无限增殖，并产生各种细胞类型。在再生过程中，临时细胞外基质 (ECM) 向 hPSC 提供多种信号，并指导新基质形成的过程。关于 hPSC 如何感知和响应周围环境的信号的基本问题。 hPSC-ECM 相互作用研究面临的主要障碍在于使用来自解剖集落的细胞团的 hPSC 培养技术的限制。在产生所需的组织特异性细胞类型之前，经过自发分化到所有三个胚胎胚层的阶段。本研究的目标是生成并表征单层单细胞培养模型，以便研究生物现象。我们将评估单层培养的 hPSC 进行上皮间质转化 (EMT) 并分化为特定胚层的潜力，并利用单细胞单层培养方法探索其作用。对称/不对称细胞分裂对 hPSC 多能性和谱系规范的调节 这将通过采用微接触印刷技术在单细胞水平上控制细胞极性和细胞骨架组织来实现。 (2) 确定单层培养的 hPSC 进行 EMT 的潜力，以及 (2) 确定不对称分裂在 hPSC 命运规范中的作用。本研究开发的新方法在许多研究领域具有广泛的适用性。包括组织工程和再生医学、疾病建模和药物测试。