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

基本信息

批准号：
9302677
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
项目类别：
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/9302677
关键词：
Address Adherent Culture Alpha Cell Area Biological Phenomena Cell Culture Techniques Cell Differentiation process Cell Lineage Cell Polarity Cell Shape Cell division Cells Cues Cultured Cells Cytoskeletal Modeling Daughter Disease Disease model Embryo Engineering Engraftment Environment Extracellular Matrix Generations Germ Layers Goals Homing Investigation Maps Mechanics Medical Methods Mitotic Mitotic spindle Modeling Musculoskeletal Diseases Natural regeneration Pattern Physiological Printing Process Proliferating Proteins Regenerative Medicine Regulation Reproducibility Research Role Shapes Signal Transduction Stem cells Techniques Testing Tissue Engineering Tissues Trauma Vision Work cell type daughter cell drug testing epithelial to mesenchymal transition human pluripotent stem cell in vivo innovation insight monolayer novel novel strategies pluripotency polarized cell prospective public health relevance regenerative therapy self-renewal stem cell fate specification tool

项目摘要

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（由适用提供）：由于创伤和各种肌肉骨骼疾病而引起的再生策略非常需要再生策略。人类多能干细胞（HPSC）为再生疗法提供了有前途的工具，因为它们具有独特的自我更新能力，几乎无限期地增殖，并升至各种细胞类型。在再生过程中，临时细胞外基质（ECM）为HPSC提供了多个信号，并指导新矩阵形成的过程。关于HPSC如何感知和应对周围环境的线索的基本问题，等待进一步的探索。 HPSC-ECM相互作用的研究面临的主要障碍在于限制HPSCS培养技术，这些技术使用了从解剖菌落中使用细胞的夹具。在这些培养方法中，HPSC不可避免地经过了赞助分化的阶段，到达所需组织特异性细胞类型之前的所有三个胚胎菌层。这项研究的目的是生成和表征单层单细胞培养模型，该模型允许与HPSC谱系规范相关的生物学现象的投资。我们将评估单层中培养的HPSC进行上皮到间质转变（EMT）的潜力，并与特定的细菌层分化。使用单细胞单层培养方法，我们将探讨对称/非对称细胞分裂在调节HPSC多能和谱系规范中的作用。这将通过采用微接合印刷技术来控制单细胞水平的细胞极性和细胞骨架组织来实现。该项目具有特定的目的：（1）确定单层中培养的HPSC进行EMT的潜力，（2）确定不对称分裂在HPSC命运规范中的作用。这项研究中开发的新方法对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。