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

间充质基质细胞 (MSC) 是在体内大多数组织中发现的稀有细胞群，对于维持这些结构具有不可估量的作用，除了形成骨骼、肌肉或脂肪组织外，MSC 还可以起到免疫抑制作用。通过调节免疫细胞（ImC）来消除炎症，并通过多种机制促进组织修复，例如释放可溶性因子或脂质结合囊泡 - 细胞外囊泡（EV）。然而，作为基于细胞的疗法进行试验时，需要大量的 EV 可能会减少这种细胞需求，但患者治疗本身就需要大量的细胞，因此在输注细胞或 EV 之前扩增 MSC 是必不可少的。间充质干细胞在实验室培养中随着时间的推移自发分化，失去其关键的幼稚免疫调节（ImM）能力。此外，由于来自老年供体的间充质干细胞的效力相对较年轻供体的间充质干细胞的效力较低，这限制了间充质干细胞的数量。目前可以接受这种疗法治疗的患者数量以及培养保留其重要抗炎和组织修复能力的功能性原始 MSC 的能力仍然是科学研究的关键目标，因此，需要研究新的方法来扩展。在实验室中保持 MSC 处于初始状态以进行治疗。之前的观察表明，MSC 的生长条件可能会发生改变，从而影响其代谢，但仍保留其 ImM 和修复特性。例如，MSC 粘附的变化会影响其能量。 MSC 细胞培养中补充小分子也可以重现这种效应并维持这些细胞的 ImM 功能，这代表了培养个体患者治疗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 治疗抗炎潜力仍有待确定，因此，使用替代生长条件可能允许扩增治疗相关的 MSC 及其 EV。潜力虽然活跃生长的细胞不断释放 EV，而 EV 含有多种物质，包括可溶性因子、DNA 和其他蛋白质，但 EV 治疗炎症性疾病已被证明与毒副作用无关。该项目将建立在初步发现的基础上，探索新型培养系统产生大规模功能细胞培养物的潜力，并将通过生物反应器的扩展进行评估。基于微载体生物反应器，最初是小规模的，以优化培养条件，该系统允许 MSC 快速生长，其数量比传统的实验室培养更快，将通过测量生长、代谢概况、ImM 来评估扩展的 MSC“质量”。还将分析 MSC EV 对 ImC 的功能以及调节这些培养系统中 MSC 释放的可溶性因子的能力，这种方法可能会导致一种新的诊断测试，以筛选大规模培养物以进行治疗。除此之外，还将对从生物反应器扩增中收集的 MSC EV 进行检查和表征，并评估它们对其他 ImC 的影响。总体而言，该项目将提供新的生物处理和诊断方法，以扩大 MSC 培养的规模。产生治疗所需数量的细胞或 EV，这有可能揭示通过调节老年供体的 MSC 体外生长条件来逆转其功能下降的新策略。