ICF: Hollow-fiber bioreactor technology to explore mechanisms and delivery of cellular therapy during machine perfusion of donated human kidneys

肾脏有许多功能，包括清洁血液和清除废物。如果肾脏衰竭，肾移植几乎总是最好的治疗方法。在英国，大多数肾脏移植手术都来自已故捐赠者。摘除、运输和移植肾脏的过程需要时间，而且会造成损害。一些在潜在器官捐赠者体内工作良好的肾脏无法忍受这种损害，因此不能用于移植。这种缺乏合适的肾脏导致肾衰竭患者的等待时间很长。一些需要肾脏移植的人永远得不到肾移植。人们已经尝试了许多治疗方法来限制摘除、运输和移植这些肾脏的破坏性影响。一种选择涉及一种机器，当肾脏在体外时，机器会将液体泵到肾脏周围。这就是所谓的“机器灌流”。它允许将其他治疗方法直接输送到肾脏我们的研究小组已经证明，使用多潜能成体祖细胞或MAPC®细胞的细胞疗法可以在机器灌流期间输送到肾脏。MAPC细胞取自人的骨髓并在实验室中生长，通过其动脉被输送到肾脏。它们似乎改善了肾脏的质量。然而，有两个主要障碍阻碍了我们使用这些肾脏进行移植。首先，对MAPC电池在这种情况下是如何工作的还没有很好的了解。其次，将MAPC细胞输送到肾脏的最佳方法尚不清楚。本研究项目旨在消除这些障碍。为了实现这一目标，我将使用一种新的方法来提供这种细胞治疗。MAPC细胞不会被移植到肾脏中，MAPC细胞将被保存在一个被称为“生物反应器”的塑料容器中。我花了9个月的时间寻找在这个生物反应器中培养MAPC细胞的最佳条件。我还使用了以前肾脏机器灌流的液体，并证明这会激活生物反应器中的MAPC细胞，这将使它们对肾脏产生影响。推进这一项目将涉及对人类肾脏进行机器灌流。这些肾脏将被捐赠，但随后被认为不适合移植。在最近亲属同意的情况下，我们可以在这个获得批准的研究项目中使用这份宝贵的礼物。这套肾脏在我们的实验后将不会被移植。我们将包括一个MAPC细胞的生物反应器作为机器灌流装置的一部分。MAPC细胞和肾脏可以交流，这意味着MAPC细胞可以治疗肾脏。然而，MAPC细胞和肾脏是在不同的隔室中。这为以受控方式研究肾脏和MAPC细胞之间的相互作用提供了独特的机会。重要的是，我们可以看到肾脏对MAPC细胞有什么影响。改变生物反应器的属性将允许我修改两者的沟通和互动方式这是一种独特的实验类型。我希望这将回答几个重要的问题：MAPC细胞是如何激活的？生物反应器中的MAPC细胞如何影响人类肾脏？它们是如何产生这种影响的？需要什么类型的互动才能产生效果？回答这些问题将使我们更接近于使用这种治疗方法进行肾脏移植。这可能会增加可供移植的肾脏数量，减少等待时间，并给等待名单上的患者带来希望。这项研究的潜在影响也更广泛。损害捐赠器官的过程也发生在常见的健康问题中，如心脏病发作和中风。例如，目前有临床试验将MAPC细胞作为中风的治疗方法。对这些细胞疗法是如何激活的，它们是如何产生效果的，以及它们如何被提供的新见解可能适用于广泛的环境。因此，该项目的发现可能导致对更广泛的疾病进行更好的治疗。

Kidneys have many functions, including cleaning the blood and removing waste. If the kidneys fail a kidney transplant is almost always the best treatment. Most kidney transplants in the UK come from deceased donors. The process of removing, transporting, and transplanting a kidney takes time and causes damage. Some kidneys which are working well in the potential organ donor cannot tolerate this damage, and therefore are not used for transplant. This lack of suitable kidneys leads to long waiting times for people with kidney failure. Some people who need a kidney transplant never get oneMany treatments have been tried to limit the damaging effects of removing, transporting, and transplanting these kidneys. One option involves a machine which pumps fluid around the kidney whilst it is outside of the body. This is called 'machine perfusion'. It allows the delivery of other treatments directly to a kidneyOur research group has shown that a cell therapy using Multipotent Adult Progenitor Cells or MAPC® cells can be delivered to a kidney during machine perfusion. The MAPC cells, taken from human bone marrow and grown in a laboratory, were delivered into the kidney through its artery. They appeared to improve the quality of the kidneys. However, there are two main barriers preventing us using these kidneys for transplant. Firstly, it is not well understood exactly how the MAPC cells work in this setting. Secondly, the best method for delivering the MAPC cells to the kidney is not known. This research project aims to remove those barriersTo achieve this aim, I will use a new method of delivering this cell treatment. Instead of putting the MAPC cells into the kidney, the MAPC cells will be maintained in a plastic container called a 'bioreactor'. I have spent 9 months finding the best conditions to grow MAPC cells in this bioreactor. I have also used fluid from previous kidney machine perfusions and shown that this 'activates' MAPC cells in the bioreactor, which will enable them to have an impact on the kidneyMoving this project forward will involve performing machine perfusion on pairs of human kidneys. These kidneys will have been donated, but then deemed unsuitable for transplantation. We can use this valuable gift in this approved research project where the next of kin gives consent. This set of kidneys will not be transplanted after our experimentsWe will include a bioreactor of MAPC cells as part of the machine perfusion device. The MAPC cells and the kidney can communicate, meaning the MAPC cells can treat the kidney. However, the MAPC cells and the kidney are in separate compartments. This gives the unique opportunity to study the interaction between the kidney and the MAPC cells in a controlled way. Importantly, we can see what effects the kidney has on the MAPC cells. Changing the properties of the bioreactor will allow me to modify how the two communicate and interactThis is a unique type of experiment. I hope that this will answer several important questions: How are MAPC cells activated? How do MAPC cells in a bioreactor affect a human kidney? How do they have this effect? What types of interaction are needed for them to have their effect? Answering these questions will bring us closer to transplanting kidneys using this treatment. This could increase the number of kidneys available for transplant, reducing waiting times, and giving hope to patients on the wait listThe potential implications of this research are also more wide reaching. The processes which injure donated organs also happen in common health problems like heart attacks and strokes. For example, there are currently clinical trials looking at MAPC cells as a treatment for strokes. New insights into how these cell therapies are activated, how they have their effect, and how they can be delivered could be applicable in a wide range of settings. Findings from this project could therefore lead to improved treatments for a much wider range of diseases