Enhancing the potency of mesenchymal stem cell therapies for kidney diseases using lab-on-a-particle technology

间充质干细胞(MSCs)是一种很有前途的治疗多种其他疾病的方法。 难治性疾病，很大程度上是由于它们产生的旁分泌因子调节 炎症，促进血管生成，抑制细胞凋亡。基于MSC的疗法正在成为 探索治疗多种肾脏和泌尿系统疾病，包括尿路 大小便失禁、狼疮性肾炎和急性肾损伤转变为慢性肾脏疾病。 不幸的是，翻译研究中不同的结果阻碍了MOST的进展 MSC疗法通过了早期临床试验。也许翻译的最大障碍是 MSCs分泌功能的固有异质性，已被证明根据不同的 用于细胞扩增的初始组织来源和条件。目前，MSCs是 通过与细胞干细胞相关但与细胞干细胞无关的表面蛋白鉴定 它们在治疗上的重要分泌功能，进一步加剧了临床上的差异 翻译。技术丰富了我们对干细胞之间直接关系的理解 分泌功能和再生潜力将被证明是现有的标准化的关键 治疗和设计更有效的细胞疗法来治疗这些慢性和破坏性的疾病 疾病。虽然用于治疗效力的功能分析方法正在逐渐被 集成到MSC开发管道中，目前还没有能够快速 基于治疗重要分泌物的单个MSC克隆的检测和丰富 各种因素。我们提出了一种基于粒子平台的新型实验室的开发，它允许快速 利用结构化的空腔将单个MSCs分离为水凝胶微粒 用于细胞黏附的固体基板和用于形成均匀微滴的模板。这种方法 将能够同时分析细胞表面和分泌的蛋白质，并允许回收 使用标准的荧光激活细胞分选器(FAC)对所需克隆进行分类。我们将评估 根据分泌血管内皮生长因子筛选克隆并鉴定其特性 慢性肾损伤患者急性肾损伤相关组织再生功能的体外检测 肾脏疾病过渡。我们的新技术承诺消除这一重大障碍 功能干细胞选择，以推动下一代MSC治疗肾脏和 泌尿系统疾病。

Mesenchymal stem cells (MSCs) are a promising treatment modality for a multitude of otherwise intractable diseases, largely due to their production of paracrine factors which modulate inflammation, promote angiogenesis, and inhibit apoptosis. MSC-based therapies are being explored for the treatment of numerous kidney and urological diseases including urinary incontinence, lupus nephritis, and acute kidney injury transitioning to chronic kidney disease. Unfortunately, disparate results in translational studies have hindered the progression of most MSC therapies past early stage clinical trials. Perhaps the greatest barrier to translation is the inherent heterogeneity in secretory function of MSCs, which has been shown to vary based on both the initial tissue source and conditions used for cell expansion. Currently, MSCs are identified through surface proteins which correlate to cell stemness but are disconnected from their therapeutically important secretory functions, further exacerbating differences in clinical translation. Technologies enriching our understanding of the direct relation between stem cell secretory function and regenerative potential will prove crucial for the standardization of existing treatments and engineering more effective cell therapies for these chronic and devastating diseases. While functional profiling approaches for therapeutic potency are gradually being integrated into MSC development pipelines, there are currently no technologies capable of rapidly detecting and enriching individual MSC clones based on therapeutically important secreted factors. We propose the development of a novel lab on a particle platform, which allows the rapid isolation of individual MSCs into hydrogel microparticles with a structured cavity that provides a solid substrate for cell adhesion and a template for uniform microdroplet formation. This approach will enable profiling of both cell surface and secreted proteins simultaneously, and allow recovery of desired clones using standard fluorescence-activated cell sorters (FACS). We will evaluate the clones selected based on secretion of vascular endothelial growth factor and characterize their function in vitro assays relevant to tissue regeneration relevant to acute kidney injury to chronic kidney disease transition. Our new technology promises to remove this significant barrier in functional stem cell selection to drive the next-generation of MSC therapies for kidney and urologic diseases.