Decellularized Stem Cell Matrix Rejuvenates Human Cells from Herniated Discs

脱细胞干细胞基质使椎间盘突出的人体细胞恢复活力

• 批准号: 8856503
• 负责人: Ming Pei
• 金额: $ 7.4万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8856503
• 关键词: Adult; Age; Autologous; Biochemical; Biological; Candidate Disease Gene; Cartilage; Cell Therapy; Cells; Chondrocyte-like Cell; Clinical; Clinical Treatment; Collagen; Collagen Gene; DNA; Deposition; ENG gene; Extracellular Matrix; Family suidae; Focus Groups; GAG Gene; Galactosidase; Gene Expression; Genes; Goals; Health; Histology; Human; Hypoxia; Immunohistochemistry; In Vitro; Incubators; Inorganic Sulfates; Intervertebral disc structure; Lead; Low Back Pain; Methods; Microarray Analysis; Molecular Profiling; Ontology; Pathway interactions; Pattern; Phenotype; Physiological; Plastics; Property; Proteoglycan; RNA; Rejuvenation; Selection Criteria; Serum; Slipped Disk; Staining method; Stains; Stem cells; Surface; Synovial Membrane; System; Therapeutic; Time; Tissues; Unspecified or Sulfate Ion Sulfates; Validation; Water; Western Blotting; aggrecan; base; candidate selection; cell dedifferentiation; disc regeneration; fetal; fetal stem cell; flasks; intervertebral disk degeneration; minimally invasive; monolayer; novel; nucleus pulposus; older patient; reconstruction; tissue regeneration

DESCRIPTION (provided by applicant): Low back pain is often related to intervertebral disc (IVD) degeneration that results from a progressive loss of proteoglycans and water content in the nucleus pulposus (NP). Autologous disc cell-based therapy is a promising approach for IVD regeneration. Unfortunately, the current in vitro expansion of NP cells in monolayer results in dedifferentiation of these cells. Synovium-derived stem cells (SDSCs) are tissue-specific stem cells for cartilage-like tissue regeneration. Our previous study indicates that SDSC-derived extracellular matrix (ECM) can provide a microenvironment for SDSC rejuvenation by enhancing proliferation capacity and chondrogenic differentiation potential. Since NP cells are chondrocyte-like cells, it is speculated that a more tissue-specific 3D microenvironment can maintain the phenotype of seeded NP cells during ex vivo expansion instead of standard 2D culture. Our most recent study suggests that such an in vitro 3D microenvironment benefits porcine NP cells by allowing them to expand more efficiently while maintaining their differentiation phenotypes. In this study, we hypothesize that decellularized matrix deposited by human SDSCs (hSDSCs) from fetal donors can serve as a tissue-specific high-quality microenvironment for ex vivo expansion of human NP cells from elderly patients while maintaining NP cell differentiated phenotype and re-differentiation potential. Two specific aims will be performed: (1) comparison of expansion efficiency of human NP cells from herniated discs when grown on ECM deposited by hSDSCs from either fetal or adult donors; and (2) exploration of the underlying mechanism by evaluating global gene expression in human NP cells expanded on ECM from either fetal or adult donors using microarray analysis. Our long-term goal is to develop an autologous disc cell-based minimally invasive therapeutic approach toward physiological reconstruction of a biologically functional disc in a clinical setting.

描述（由申请人提供）：腰痛通常与椎间盘（IVD）退变有关，椎间盘退变是由髓核（NP）中蛋白多糖和水含量的进行性损失引起的。基于自体椎间盘细胞的治疗是一种有前途的IVD再生方法。不幸的是，目前在体外扩增的NP细胞在单层的结果，这些细胞的去分化。滑膜源性干细胞（SDSC）是用于软骨样组织再生的组织特异性干细胞。我们以前的研究表明，SDSC衍生的细胞外基质（ECM）可以通过增强增殖能力和软骨分化潜力为SDSC的年轻化提供微环境。由于NP细胞是软骨细胞样细胞，因此推测更具有组织特异性的3D微环境可以在离体扩增而不是标准2D培养期间保持接种的NP细胞的表型。我们最近的研究表明，这种体外3D微环境有利于猪NP细胞，使它们能够更有效地扩增，同时保持其分化表型。在这项研究中，我们假设由来自胎儿供体的人SDSC（hSDSC）沉积的脱细胞基质可以作为用于来自老年患者的人NP细胞的离体扩增的组织特异性高质量微环境，同时保持NP细胞分化表型和再分化潜力。将进行两个具体的目的：（1）当在由来自胎儿或成人供体的hSDSC沉积的ECM上生长时，比较来自椎间盘突出的人NP细胞的扩增效率;和（2）通过使用微阵列分析评估在来自胎儿或成人供体的ECM上扩增的人NP细胞中的整体基因表达来探索潜在机制。我们的长期目标是开发一种基于自体椎间盘细胞的微创治疗方法，在临床环境中对生物功能性椎间盘进行生理重建。