GMP-based CD133(+) cells isolation maintains progenitor angiogenic properties and enhances standardization in cardiovascular cell therapy.
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.
Gaipa
Giuseppe
G
Laboratorio Interdipartimentale di Terapia Cellulare Stefano Verri, Azienda Ospedaliera San Gerardo, Monza, Milan, Italy.
Tilenni
Manuela
M
Straino
Stefania
S
Burba
Ilaria
I
Zaccagnini
Germana
G
Belotti
Daniela
D
Biagi
Ettore
E
Valentini
Marco
M
Perseghin
Paolo
P
Parma
Matteo
M
Campli
Cristiana Di
CD
Biondi
Andrea
A
Capogrossi
Maurizio C
MC
Pompilio
Giulio
G
Pesce
Maurizio
M
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
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