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（申请人提供）：组织工程与干细胞结合是再生医学的一个新兴领域，在修复因疾病、衰老和创伤而受损的组织和器官方面具有前景。为了改善干细胞的治疗用途，研究如何提高干细胞增殖和分化的定性和定量程度，从而增加细胞生长成所需组织或结构类型的机会是很重要的。一般来说，温度对生长过程的影响是仅次于供氧的重要物理因素。更具体地说，体内研究表明，局部透热可以刺激骨形成。此外，热休克蛋白（HSPs）的上调及其在胚胎发育中的重要作用已被广泛认识。局部或全身温度升高是组织损伤后炎症期间常见的，可能在组织再生中起重要作用。然而，胚胎发育过程中温度诱导骨细胞分化和热休克蛋白过表达的机制尚不清楚。升高的温度是否会影响成体干细胞的募集和分化也是未知的。虽然对其他因素的影响已经进行了大量的研究，但对热效应对干细胞分化的影响的研究很少。因此，PI建议对热休克对间充质干细胞（mesenchal stem cell， MSC）增殖和分化的影响进行全面研究，以验证我们的假设-温和的温度升高可能促进MSC的分化和成熟。这个为期两年的试点项目的具体目标是：(1)使用合成和自组装肽水凝胶- PuraMatrix建立优化的人间充质干细胞（hMSC）增殖的3D培养条件，并评估hMSC在3D PuraMatrix培养下与传统的2D培养成骨或3D颗粒培养软骨形成相比的分化情况，使用经典的分化诱导剂；(2)在优化的三维培养环境下，探索动态热休克对hMSC增殖和向成骨细胞和软骨细胞分化的影响，并利用靶向HSP70和HSP27的sirna，研究热休克蛋白（HSPs）在热休克对hMSC分化的作用。热休克对hMSC分化为骨和软骨细胞的影响的初步研究的预期意义是：(a)促进hMSC增殖和分化为成骨细胞/软骨细胞；(b)改良成熟细胞，使已分化的细胞能像成体细胞一样有效地发挥功能；(c)提供3D MSC体外培养模型，最大限度地模拟体内分化环境；(d)为设计完全分化干细胞在生物人工器官/细胞移植中临床应用的热处理方案提供基础科学数据。