Manufacture scale up of human pericyte progenitor cells for regenerative medicine

再生干细胞疗法为治疗心血管疾病带来了新的希望。这种方法的潜力是巨大的，但还没有完全开发出来。到目前为止，心肌梗死(MI)患者的细胞治疗临床试验研究的选择相对较少，主要集中在血液和骨髓来源细胞的使用上。许多其他细胞值得关注，因为它们可能为心肌和血管重建提供一种特殊的手段。其中，血管壁细胞和周细胞受到越来越多的关注。以周细胞为基础的治疗的基本原理是稳定梗塞周围的血管，否则这些血管仍然容易泄漏和破裂。我们开发了一种从冠心病患者的人隐静脉中分离和扩增周细胞的方法。静脉是使用冠状动脉搭桥手术中遗留下来的东西获得的，不会对患者造成伤害。同样，可以通过微创手术获得一小块静脉，以分离和扩大培养中的周细胞，并获得数百万准备用于临床的细胞。我们证明了人周细胞移植在肢体缺血和心肌梗死模型中发挥了长期的治疗作用。值得注意的是，周细胞移植在梗死周围血管形成中发挥作用，并发挥一系列修复作用，包括吸引驻留的心脏干细胞和循环中的血管生成细胞。此外，周细胞可提高心肌细胞的存活率，从而减少梗塞面积。没有安全问题的报道。目前的分离/扩增方法是在一个普通的组织培养实验室中开发的。因此，我们现在需要将其升级到患者临床使用所需的标准。本研究旨在通过一系列调节程序和质量控制来开发临床级别的细胞产品，以准备首次在冠心病患者中使用自体周细胞进行临床研究，该研究对药物无反应且不适合血管重建。术语自体是指从患者的静脉中提取周细胞，扩增并重新注入患者的心脏。使用患者自己的细胞可以避免他们的排斥反应和传染病的传播，这是使用来自不同捐赠者的细胞时常见的问题。该项目包括3个工作包(WP)、6个里程碑(M)和一个预防、故障排除和替代解决方案应急计划。WP1的目标是扩大目前的方案，以提高可用于临床的细胞产量。WP2的目标是将该协议转移到国家卫生服务-血液和移植(NHSBT)的设施中，用于临床级至少6个细胞系的生产。随后将对细胞稳定性进行质量控制。WP3的目标是使用体外功能分析和心肌梗死模型来测试细胞产品的性能。该项目将通过布里斯托尔大学和NHSBT的联系，导致一种新的再生医学细胞产品的临床生产，这是英国自体细胞疗法临床应用的推荐渠道。因此，成功完成拟议的工作将使细胞产品立即可用于医疗保健系统，以造福患者。

Regenerative stem cell therapy offers new hope for treating cardiovascular disease. The potential of this approach is immense but not completely developed. Clinical trials of cell therapy in patients with myocardial infarction (MI) have so far investigated a relatively small number of options, focusing mainly on the use of blood and bone marrow-derived cells. A host of other cells merit attention as they may offer a specialised means for myocardial and vascular reconstitution. Among these, vascular mural cells and pericytes are gaining attention. The rationale of pericyte-based therapy is to stabilise peri-infarct blood vessels, which would remain otherwise prone to leak and break. We have developed a method for easy isolation and expansion of pericytes from human saphenous veins of patients with coronary artery disease. The vein is obtained without harm to the patient using the leftover from coronary artery bypass surgery. Likewise, a small piece of vein could be obtained through minimal invasive surgery to isolate and expand pericytes in culture and obtain million cells ready for clinical use. We demonstrated that transplantation of human pericytes exerts long-term therapeutic benefit in models of limb ischaemia and MI. Noteworthy, pericytes engraft in peri-infarct vascularisation and exert a spectrum of reparative actions, including attraction of resident cardiac stem cells and circulating angiogenic cells. Furthermore, pericytes improve the survival of cardiac cells thus leading to reduction of the infarct size. No safety concerns were reported. The current isolation/expansion method was developed in a common tissue culture laboratory. Hence, we now need to upgrade it to the standards required for clinical use in patients. The present study aims to develop a clinical grade cell product through a series of regulated procedures and quality controls in preparation of a first-in-man clinical study with autologous pericytes in coronary artery disease patients unresponsive to medications and not amenable to revascularization. The term autologous means that pericytes are extracted from the patient vein, expanded and reintroduced into the patient's heart. Use of patient's own cells avoids their rejection and transmission of infectious disease, which are common problems when using cells from a different donor. The project comprises 3 work-packages (WP), 6 milestones (M) and a contingency plan for prevention, troubleshooting and alternative solutions. The goal of WP1 is to scale up current protocol to improve the yield of cells available for clinical use. The goal of WP2 is to transfer the protocol to the facilities of the National Health Service-Blood and Transplant (NHSBT) for clinical-grade manufacture of at least 6 cell lines. This will be followed by quality control of cell stability. The goal of WP3 is to test the performance of the cell product using in vitro functional assays and models of MI. The project will lead to the clinical manufacture of a novel regenerative medicine cell product through the liaison of the University of Bristol and NHSBT, which is the recommended channel for clinical application of autologous cell therapy in UK. Hence, successful completion of proposed work will make the cell product available immediately to the healthcare system for patient benefit.