Investigating bioengineering approaches to produce immuno-modulatory mesenchymal stromal cells and their extracellular vesicle

研究生产免疫调节间充质基质细胞及其细胞外囊泡的生物工程方法

• 批准号: 2608627
• 金额: --
• 依托单位: Aston University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2025
• 资助国家: 英国
• 起止时间: 2025 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2608627
• 关键词: Investigating; bioengineering; approaches; produce; immuno

Mesenchymal stromal cells (MSCs) are a rare population of cells found in most tissues within the body and are invaluable in the maintenance of these structures. As well as forming bone, muscle or fatty tissues, MSCs can also be immunosuppressive. They can both resolve inflammation by modulating immune cells (ImCs) and promote tissue repair through multiple mechanisms such as the release of soluble factors or lipid-bound vesicles - extracellular vesicles (EVs). Currently, MSCs are being trialed as a cell-based therapy, however, large numbers are required. EVs may reduce this cell requirement, but are themselves needed in high quantities for patient therapy, therefore expansion of MSCs is essential before transfusion of either cells or EVs. Unfortunately, MSCs spontaneously differentiate over time in laboratory culture, losing their critical naïve immunomodulatory (ImM) abilities. Moreover, the clinical demand for MSCs remains unmet as MSCs from older donors are comparatively less potent than those from younger donors. This limits the number of patients that can be currently treated with this therapy and the ability to grow functional, naïve MSCs which retain their important anti-inflammatory and tissue repair abilities remains a key goal for scientific research. Thus, there is a need to investigate new methodologies to expand MSCs in the laboratory whilst maintaining them in a naïve state for therapy.Previous observations have revealed the potential for altering the growth conditions of MSCs which affect their metabolism yet retaining their ImM and repair properties. Changes in MSC adhesion for example, affect their energy production; a key feature of how MSCs modulate their physiology. Supplementation of small molecules to MSC cell culture can also reproduce this effect and maintain the ImM functions of these cells. These represent new approaches to culturing the large number of cells required for individual patient therapy or EV collection. How these culture methods affect MSC ImM properties or EV therapeutic anti-inflammatory potential remains to be defined. Therefore using alternative growth conditions potentially allows expansion of therapeutically relevant MSC and their EVs. The delivery of molecules from EVs to cells may have the potential to control the immune system for therapeutic benefit. While actively growing cells continuously shed EVs - and EVs contain a variety of cargos including soluble factors, DNA and other proteins - EV treatment of inflammatory conditions has been demonstrated not to be associated with toxic side effects of standard drug treatments. This project will build upon initial findings, exploring the potential of novel culture systems to generate large scale cultures of functional cells for therapy. Combining polymer-based growth surfaces with expansion in bioreactors will be evaluated. MSCs will be expanded via a microcarrier-based bioreactor, initially on a small-scale to optimise culture conditions. This system allows the rapid growth of MSCs, increasing their numbers more quickly than traditional laboratory-based culture. Expanded MSC 'quality' will be evaluated by measuring growth, metabolism profiles, ImM function on ImCs, and the MSC EV's ability to modulate inflammation. Released soluble factors from MSCs within these culture systems will also be analysed. This approach may lead to a new diagnostic test to screen large scale cultures for therapy to provide patients with optimal, functional cells. Alongside this, MSC EVs collected from bioreactor expansion will be examined and characterised, with their effects on other ImCs evaluated.Overall, this project will inform on new bioprocessing and diagnostic approaches to allow the upscaling of MSC cultures to generate the required numbers of cells or EVs required for therapy. This has the potential to reveal new strategies for reversing the reduced functionality of MSCs from older donors through modulating their in-vitro growth condition

间充质间质细胞（MSCs）是一种罕见的细胞群，存在于人体大多数组织中，在维持这些组织结构方面具有不可估量的价值。除了形成骨骼、肌肉或脂肪组织外，间充质干细胞还可以抑制免疫。它们既可以通过调节免疫细胞（ImCs）来缓解炎症，也可以通过释放可溶性因子或脂质结合囊泡-细胞外囊泡（EVs）等多种机制促进组织修复。目前，间充质干细胞正在作为一种基于细胞的疗法进行试验，然而，需要大量的干细胞。EVs可能会减少这种细胞需求，但患者治疗本身需要大量的MSCs，因此在输入细胞或EVs之前，MSCs的扩增是必不可少的。不幸的是，随着时间的推移，MSCs在实验室培养中会自发分化，失去其关键的naïve免疫调节（ImM）能力。此外，由于老年供者的骨髓间充质干细胞的效力相对低于年轻供者，临床对骨髓间充质干细胞的需求仍未得到满足。这限制了目前可以用这种疗法治疗的患者数量和培养功能性的naïve间充质干细胞的能力，这些干细胞保留了重要的抗炎和组织修复能力，仍然是科学研究的关键目标。因此，有必要研究新的方法，在实验室中扩大间充质干细胞，同时保持它们在naïve状态进行治疗。先前的观察已经揭示了改变间充质干细胞生长条件的潜力，这影响了它们的代谢，但保留了它们的ImM和修复特性。例如，MSC粘附的变化会影响它们的能量产生；这是间充质干细胞调节其生理的关键特征。在MSC细胞培养中补充小分子也可以复制这种效果，并维持这些细胞的ImM功能。这些代表了培养个体患者治疗或EV收集所需的大量细胞的新方法。这些培养方法如何影响MSC ImM特性或EV治疗性抗炎潜力仍有待确定。因此，使用其他生长条件可能会扩大治疗相关的MSC及其EVs。从ev向细胞输送分子可能具有控制免疫系统以获得治疗益处的潜力。虽然活跃生长的细胞不断释放EV - EV含有多种货物，包括可溶性因子，DNA和其他蛋白质- EV治疗炎症条件已被证明与标准药物治疗的毒副作用无关。该项目将建立在初步发现的基础上，探索新型培养系统的潜力，以产生用于治疗的功能细胞的大规模培养。将聚合物基生长表面与生物反应器中的膨胀相结合进行评估。msc将通过基于微载体的生物反应器进行扩增，最初是小规模的，以优化培养条件。该系统允许间充质干细胞快速生长，比传统的实验室培养更快地增加其数量。扩大的MSC“质量”将通过测量生长、代谢特征、ImM在imc上的功能以及MSC EV调节炎症的能力来评估。还将分析这些培养系统中MSCs释放的可溶性因子。这种方法可能会导致一种新的诊断测试，以筛选大规模培养的治疗，为患者提供最佳的功能细胞。除此之外，将对从生物反应器扩展中收集的MSC ev进行检查和表征，并评估其对其他ImCs的影响。总体而言，该项目将为新的生物处理和诊断方法提供信息，以扩大MSC培养的规模，以产生治疗所需的细胞或ev数量。这有可能揭示通过调节老年供体间充质干细胞的体外生长条件来逆转其功能下降的新策略