title: GMP-conformant on-site manufacturing of a CD133+stem cell product for cardiovascular regeneration.
abstract: 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 × 106viable CD133+cells with a mean log10depletion 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.
authors.affiliation: 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.
authors.forename: Anna
authors.initials: A
authors.lastname: Skorska
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Paula
authors.initials: P
authors.lastname: Müller
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Gaebel
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Große
authors.affiliation: 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.
authors.forename: Heiko
authors.initials: H
authors.lastname: Lemcke
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Cornelia A
authors.initials: CA
authors.lastname: Lux
authors.affiliation: Institute for Clinical Chemistry and Laboratory Medicine (ILAB), Rostock University Medical Center, Ernst-Heydemann-Straße 6, Rostock, 18057, Germany.
authors.forename: Manuela
authors.initials: M
authors.lastname: Bastian
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Frauke
authors.initials: F
authors.lastname: Hausburg
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Zarniko
authors.affiliation: Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 35, Rostock, 18057, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Bubritzki
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Ruch
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 68, Rostock, 18057, Germany.
authors.forename: Gudrun
authors.initials: G
authors.lastname: Tiedemann
authors.affiliation: 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.
authors.forename: Robert
authors.initials: R
authors.lastname: David
authors.affiliation: Department Life, Light and Matter of the Interdisciplinary Faculty at Rostock University, Albert-Einstein Straße 25, Rostock, 18059, Germany. gustav.steinhoff@med.uni-rostock.de.|Department of Cardiac Surgery, Rostock University Medical Center, Schillingallee 35, Rostock, 18057, Germany. gustav.steinhoff@med.uni-rostock.de.
authors.forename: Gustav
authors.initials: G
authors.lastname: Steinhoff
mesh_terms: D000071916:AC133 Antigen / Q000235:genetics / Q000378:metabolism; D000368:Aged; D000818:Animals; D057205:Automation, Laboratory / Q000295:instrumentation*; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D004195:Disease Models, Animal; D005260:Female; D015870:Gene Expression; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D008297:Male; D051379:Mice; D016513:Mice, SCID; D009203:Myocardial Infarction / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D020127:Recovery of Function / Q000502:physiology; D012038:Regeneration / Q000502:physiology*; D014019:Tissue Donors
chemical_list: D000071916:AC133 Antigen; D015415:Biomarkers; C000606024:Prom1 protein, mouse
keywords: Adult hematopoietic stem cells; Advanced therapy medicinal product (ATMP); CD133+ cells; Cardiovascular regeneration; Clinical translation; Good Manufacturing Practice (GMP); Prodigy; Stem cell transplantation