Heat Shock Effects on Human Mesenchymal Stem Cell Differentiation in Hydrogel

热休克对水凝胶中人间充质干细胞分化的影响

• 批准号: 7693161
• 负责人: Sihong Wang
• 金额: $ 14.32万
• 依托单位: CITY COLLEGE OF NEW YORK
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7693161
• 关键词: Adult; Affect; Age; Aging; Ascorbic Acid; Award; Bioartificial Organs; Biocompatible Materials; Biomedical Engineering; Bioreactors; Bone Marrow; Calcium; Cartilage; Cell Culture Techniques; Cell Differentiation process; Cell Maturation; Cell Proliferation; Cell Transplantation; Cell model; Cells; Chondrocytes; Chondrogenesis; Cities; Clinical; Coculture Techniques; Collagen Type I; Collagen Type II; Coupled; Data; Defect; Deposition; Development; Devices; Dexamethasone; Diathermy; Differentiation Inducer; Disease; Embryonic Development; Environment; Evaluation; Family; Gel; Generations; Genes; Glycerophosphates; Goals; Growth; Growth Factor; HSPB1 gene; Heat shock proteins; Heat-Shock Proteins 70; Heat-Shock Response; Heating; Histocompatibility Testing; Hour; Human; Hydrogels; In Vitro; Inflammation; Inflammatory; Injection of therapeutic agent; Injury; Investigation; Knowledge; Lead; Location; Mechanics; Mesenchymal; Mesenchymal Stem Cells; Modeling; Molecular; New York; New York City; Organ; Osteoblasts; Osteogenesis; Oxygen; Oxygen measurement, partial pressure, arterial; Peptides; Perfusion; Pilot Projects; Polymerase Chain Reaction; Process; Protein Overexpression; Proteolysis; Protocols documentation; RNA Interference; Regenerative Medicine; Research; Reverse Transcriptase Polymerase Chain Reaction; Reverse Transcription; Role; SOX9 protein; Staining method; Stains; Stem Cell Research; Stem cells; Stromal Cells; Structure; Surface; Technology; Temperature; Therapeutic; Time; Tissue Engineering; Tissues; Transplantation; Trauma; Treatment Protocols; Up-Regulation; adult stem cell; aged; aggrecan; bone; bone cell; cartilage cell; cell motility; cell type; college; cytokine; design; experience; improved; in vivo; injured; inorganic phosphate; knock-down; migration; osteogenic; overexpression; public health relevance; repaired; restoration; stem cell differentiation; tissue regeneration; transcription factor

DESCRIPTION (provided by applicant): Tissue engineering combined with stem cells, an emerging field of regenerative medicine, holds promise for the restoration of tissues and organs damaged by diseases, aging, and trauma. In order to improve the therapeutic usage of stem cells, it is important to investigate ways to enhance the qualitative and quantitative extent of stem cell proliferation and differentiation, and thus increase the chances for cells to grow into desired types of tissues or structures. Temperature is an important physical factor only second to oxygen supply in its influence on growth processes in general. More specifically, in vivo study has shown that bone formation is stimulated by local diathermy. In addition, the upregulation of heat shock proteins (HSPs) and their important roles in embryonic development have been widely recognized. Local or systemic temperature elevation is common during inflammation after tissue injury and may have an important role in tissue regeneration. However, the mechanisms for temperature-induced bone cell differentiation as well as overexpression of HSPs during embryonic development are unclear. Whether or not elevated temperature affects adult stem cell recruitment and differentiation is also unknown. Although intensive investigations have been conducted on the influences of other factors, few studies have been done on thermal effects on stem cell differentiation. Therefore, the PI proposes to conduct a comprehensive study on heat shock effects on mesenchymal stem cell (MSC) proliferation and differentiation to investigate our hypothesis - that mild temperature increase may facilitate MSC differentiation and maturation. The specific aims for this two-year pilot project are: (1) to establish an optimized 3D culture condition for human mesenchymal stem cell (hMSC) proliferation using a synthetic and self-assembling peptide hydrogel - PuraMatrix, and to evaluate hMSC differentiation on 3D PuraMatrix culture versus conventional 2D culture for osteogenesis or 3D pellet culture for chondrogenesis, using classic differentiation inducers; and (2) to explore dynamic heat-shock effects on hMSC proliferation and differentiation into osteoblasts and chondrocytes in the optimized 3D culture environment, and to investigate the role of heat shock proteins (HSPs) in heat shock effects on hMSC differentiation using siRNAs targeting HSP70 and HSP27. The expected significances of this pilot study of heat shock effects on hMSC differentiation into bone and cartilage cells are: (a) to facilitate hMSC proliferation and differentiation into osteoblasts/chondrocytes; (b) to improve maturature so that differentiated cells can function as efficiently as adult cells; (c) to provide a 3D in vitro MSC culture model to maximally mimic the in vivo differentiation environment; and (d) to provide basic scientific data for the design of a thermal treatment protocol that is clinically applicable using fully differentiated stem cells in bioartifical organ/cell transplantation. PUBLIC HEALTH RELEVANCE: This pilot study may lead to a new strategy that uses thermal treatments to enhance the maturature levels of osteoblasts (bone cells) and chondrocytes (cartilage cells) differentiated from mesenchymal stem cells. With increased maturature, differentiated cells are better to perform functions similar to adult cells in tissue generation for the repair of injured or aged organs. In addition, the award of this pilot proposal will help to establish long term stem cell research in the Biomedical Engineering (BME) Department at the City College of New York (CCNY).

描述（由申请人提供）：组织工程与干细胞结合，新兴的再生医学领域，有望恢复受疾病，衰老和创伤受损的组织和器官。为了改善干细胞的治疗用法，重要的是要研究增强干细胞增殖和分化的定性和定量程度的方法，从而增加了细胞成长为所需的组织或结构类型的机会。温度是重要的物理因素，仅在其对生长过程的影响方面仅次于氧气供应。更具体地说，体内研究表明，骨形成是由局部核心刺激的。此外，热休克蛋白（HSP）的上调及其在胚胎发育中的重要作用已得到广泛认可。在组织损伤后炎症期间，局部或全身温度升高是常见的，并且可能在组织再生中起重要作用。但是，尚不清楚温度引起的骨细胞分化以及HSP的过表达的机制尚不清楚。温度是否升高会影响成年干细胞募集和分化。尽管已经对其他因素的影响进行了深入的研究，但对干细胞分化的热影响很少进行研究。因此，PI建议对中充质干细胞（MSC）增殖和分化的热休克作用进行全面研究，以研究我们的假设 - 轻度温度升高可能促进MSC的分化和成熟。该为期两年的试验项目的具体目的是：（1）使用合成和自组装的肽水凝胶-puramatrix建立人类间充质干细胞（HMSC）增殖的优化3D培养条件，并评估HMSC在3D Puramatrix Cultance crivisial 2D培养物上使用经典的稳定培养物进行稳定培养的HMSC差异，以实现3D Puramatrix培养物，以实现3D培养的态度，以稳定的态度培养3D pellet sogenseiss或3D peldereiss corport of 3d pellet culdeiss of 3D peldeisiss corporeiss of 3D P. （2）在优化的3D培养环境中探索动态热震影响对成骨细胞和软骨细胞的动态热冲作用，并研究热休克蛋白（HSP）（HSP）在使用SIRNAS靶向HSP70和HSP27的HMSC分化中热休克蛋白（HSP）对HMSC差异的作用。这项对热休克对HMSC分化为骨骼和软骨细胞分化的试点研究的预期意义是：（a）促进HMSC增殖并分化为成骨细胞/软骨细胞； （b）改善成熟度以使分化细胞可以像成人细胞一样有效地发挥作用； （c）提供3D体外MSC培养模型，以最大程度地模仿体内分化环境； （d）提供基本的科学数据，以设计热处理方案的设计，该方案在临床上使用生物体型器官/细胞移植中使用完全分化的干细胞。 公共卫生相关性：这项初步研究可能导致一种新策略，该策略使用热处理来增强成骨细胞（骨细胞）和软骨细胞（软骨细胞）的成熟水平，从而从间充质干细胞区分开来。随着成熟的增加，分化细胞更好地执行类似于组织产生的成年细胞的功能，以修复受伤或老化的器官。此外，该试点建议的裁决将有助于在纽约市城市学院（CCNY）的生物医学工程（BME）系中建立长期的干细胞研究。