Decellularized Stem Cell Matrix Rejuvenates Human Cells from Herniated Discs

描述（由申请人提供）：腰痛通常与椎间盘（IVD）退变有关，这是由髓核（NP）中蛋白聚糖和水含量的逐渐丧失引起的。自体椎间盘细胞为基础的治疗是一种很有前途的IVD再生方法。不幸的是，目前NP细胞在体外单层扩增导致这些细胞去分化。滑膜源性干细胞（SDSCs）是用于软骨样组织再生的组织特异性干细胞。我们之前的研究表明，SDSC衍生的细胞外基质（ECM）可以通过增强SDSC的增殖能力和软骨分化潜能，为SDSC年轻化提供微环境。由于NP细胞是软骨细胞样细胞，因此推测一个更具组织特异性的3D微环境可以在体外扩增过程中维持种子NP细胞的表型，而不是标准的2D培养。我们最近的研究表明，这种体外3D微环境有利于猪NP细胞，使它们在保持分化表型的同时更有效地扩展。在这项研究中，我们假设来自胎儿供体的人SDSCs （hSDSCs）沉积的脱细胞基质可以作为一个组织特异性的高质量微环境，用于老年患者的人NP细胞的体外扩增，同时保持NP细胞的分化表型和再分化潜力。主要有两个目的：(1)比较来自椎间盘突出的人NP细胞在由胎儿或成人供体hSDSCs沉积的ECM上的扩增效率；(2)利用微阵列分析技术，通过评估胚胎或成人供体ECM上扩增的人类NP细胞的整体基因表达，探索其潜在机制。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，用于临床环境中生物功能椎间盘的生理重建。

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.