Skorska Anna, Müller Paula, Gaebel Ralf, Große Jana, Lemcke Heiko, Lux Cornelia A, Bastian Manuela, Hausburg Frauke, Zarniko Nicole, Bubritzki Sandra, Ruch Ulrike, Tiedemann Gudrun, David Robert, Steinhoff Gustav
Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
Department Life, Light and Matter of the Interdisciplinary Faculty at Rostock University, Albert-Einstein Straße 25, Rostock, 18059, Germany.
Stem Cell Res Ther. 2017 Feb 10;8(1):33. doi: 10.1186/s13287-016-0467-0.
CD133 stem cells represent a promising subpopulation for innovative cell-based therapies in cardiovascular regeneration. Several clinical trials have shown remarkable beneficial effects following their intramyocardial transplantation. Yet, the purification of CD133 stem cells is typically performed in centralized clean room facilities using semi-automatic manufacturing processes based on magnetic cell sorting (MACS®). However, this requires time-consuming and cost-intensive logistics.
CD133 stem cells were purified from patient-derived sternal bone marrow using the recently developed automatic CliniMACS Prodigy® BM-133 System (Prodigy). The entire manufacturing process, as well as the subsequent quality control of the final cell product (CP), were realized on-site and in compliance with EU guidelines for Good Manufacturing Practice. The biological activity of automatically isolated CD133 cells was evaluated and compared to manually isolated CD133 cells via functional assays as well as immunofluorescence microscopy. In addition, the regenerative potential of purified stem cells was assessed 3 weeks after transplantation in immunodeficient mice which had been subjected to experimental myocardial infarction.
We established for the first time an on-site manufacturing procedure for stem CPs intended for the treatment of ischemic heart diseases using an automatized system. On average, 0.88 × 10 viable CD133 cells with a mean log depletion of 3.23 ± 0.19 of non-target cells were isolated. Furthermore, we demonstrated that these automatically isolated cells bear proliferation and differentiation capacities comparable to manually isolated cells in vitro. Moreover, the automatically generated CP shows equal cardiac regeneration potential in vivo.
Our results indicate that the Prodigy is a powerful system for automatic manufacturing of a CD133 CP within few hours. Compared to conventional manufacturing processes, future clinical application of this system offers multiple benefits including stable CP quality and on-site purification under reduced clean room requirements. This will allow saving of time, reduced logistics and diminished costs.
CD133干细胞是心血管再生创新细胞疗法中一个很有前景的亚群。多项临床试验表明，心肌内移植后有显著的有益效果。然而，CD133干细胞的纯化通常在集中的洁净室设施中使用基于磁性细胞分选（MACS®）的半自动制造工艺进行。但这需要耗时且成本高昂的物流。
使用最近开发的自动CliniMACS Prodigy® BM - 133系统（Prodigy）从患者来源的胸骨骨髓中纯化CD133干细胞。整个制造过程以及最终细胞产品（CP）的后续质量控制在现场完成，并符合欧盟药品生产质量管理规范指南。通过功能测定以及免疫荧光显微镜评估自动分离的CD133细胞的生物活性，并与手动分离的CD133细胞进行比较。此外，在接受实验性心肌梗死的免疫缺陷小鼠移植3周后，评估纯化干细胞的再生潜力。
我们首次使用自动化系统建立了用于治疗缺血性心脏病的干细胞CP的现场制造程序。平均分离出0.88×10个存活的CD133细胞，非靶细胞的平均对数清除率为3.23±0.19。此外，我们证明这些自动分离的细胞在体外具有与手动分离的细胞相当的增殖和分化能力。而且，自动生成的CP在体内显示出同等的心脏再生潜力。
我们的结果表明，Prodigy是一个强大的系统，可在数小时内自动制造CD133 CP。与传统制造工艺相比，该系统未来的临床应用具有多种优势，包括稳定的CP质量以及在降低洁净室要求的情况下进行现场纯化。这将节省时间、减少物流并降低成本。
