title: GMP-based CD133(+) cells isolation maintains progenitor angiogenic properties and enhances standardization in cardiovascular cell therapy.
abstract: The aim of the present study was to develop and validate a good manufacturing practice (GMP) compliant procedure for the preparation of bone marrow (BM) derived CD133(+) cells for cardiovascular repair. Starting from available laboratory protocols to purify CD133(+) cells from human cord blood, we implemented these procedures in a GMP facility and applied quality control conditions defining purity, microbiological safety and vitality of CD133(+) cells. Validation of CD133(+) cells isolation and release process were performed according to a two-step experimental program comprising release quality checking (step 1) as well as 'proofs of principle' of their phenotypic integrity and biological function (step 2). This testing program was accomplished using in vitro culture assays and in vivo testing in an immunosuppressed mouse model of hindlimb ischemia. These criteria and procedures were successfully applied to GMP production of CD133(+) cells from the BM for an ongoing clinical trial of autologous stem cells administration into patients with ischemic cardiomyopathy. Our results show that GMP implementation of currently available protocols for CD133(+) cells selection is feasible and reproducible, and enables the production of cells having a full biological potential according to the most recent quality requirements by European Regulatory Agencies.
authors.affiliation: Laboratorio Interdipartimentale di Terapia Cellulare Stefano Verri, Azienda Ospedaliera San Gerardo, Monza, Milan, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gaipa
authors.forename: Manuela
authors.initials: M
authors.lastname: Tilenni
authors.forename: Stefania
authors.initials: S
authors.lastname: Straino
authors.forename: Ilaria
authors.initials: I
authors.lastname: Burba
authors.forename: Germana
authors.initials: G
authors.lastname: Zaccagnini
authors.forename: Daniela
authors.initials: D
authors.lastname: Belotti
authors.forename: Ettore
authors.initials: E
authors.lastname: Biagi
authors.forename: Marco
authors.initials: M
authors.lastname: Valentini
authors.forename: Paolo
authors.initials: P
authors.lastname: Perseghin
authors.forename: Matteo
authors.initials: M
authors.lastname: Parma
authors.forename: Cristiana Di
authors.initials: CD
authors.lastname: Campli
authors.forename: Andrea
authors.initials: A
authors.lastname: Biondi
authors.forename: Maurizio C
authors.initials: MC
authors.lastname: Capogrossi
authors.forename: Giulio
authors.initials: G
authors.lastname: Pompilio
authors.forename: Maurizio
authors.initials: M
authors.lastname: Pesce
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D002318:Cardiovascular Diseases / Q000628:therapy*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods* / Q000592:standards*; D002470:Cell Survival; D005312:Fetal Blood / Q000166:cytology; D006023:Glycoproteins / Q000378:metabolism*; D006614:Hindlimb / Q000098:blood supply; D006801:Humans; D051379:Mice; D017202:Myocardial Ischemia / Q000473:pathology / Q000628:therapy; D018919:Neovascularization, Physiologic; D010455:Peptides / Q000378:metabolism*; D010641:Phenotype; D011786:Quality Control; D012015:Reference Standards; D033581:Stem Cell Transplantation / Q000592:standards*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D040301:Vascular Endothelial Growth Factor Receptor-2