title: Are purified or expanded cord blood-derived CD133+ cells better at improving cardiac function?
abstract: Endothelial progenitor cells (EPCs), which express the CD133 marker, can differentiate into mature endothelial cells (ECs) and create new blood vessels. Normal angiogenesis is unable to repair the injured tissues that result from myocardial infarction (MI). Patients who have high cardiovascular risks have fewer EPCs and their EPCs exhibit greater in vitro senescence. Human umbilical cord blood (HUCB)-derived EPCs could be an alternative to rescue impaired stem cell function in the sick and elderly. The aim of this study was to purify HUCB-derived CD133(+) cells, expand them in vitro and evaluate the efficacy of the purified and expanded cells in treating MI in rats. CD133(+) cells were selected for using CD133-coupled magnetic microbeads. Purified cells stained positive for EPC markers. The cells were expanded and differentiated in media supplemented with fetal calf serum and basic fibroblast growth factor, insulin-like growth factor-I and vascular endothelial growth factor (VEGF). Differentiation was confirmed by lack of staining for EPC markers. These expanded cells exhibited increased expression of mature EC markers and formed tubule-like structures in vitro. Only the expanded cells expressed VEGF mRNA. Cells were expanded up to 70-fold during 60 days of culture, and they retained their functional activity. Finally, we evaluated the therapeutic potential of purified and expanded CD133(+) cells in treating MI by intramyocardially injecting them into a rat model of MI. Rats were divided into three groups: A (purified CD133(+) cells-injected); B (expanded CD133(+) cells-injected) and C (saline buffer-injected). We observed a significant improvement in left ventricular ejection fraction for groups A and B. In summary, CD133(+) cells can be purified from HUCB, expanded in vitro without loosing their biological activity, and both purified and expanded cells show promising results for use in cellular cardiomyoplasty. However, further pre-clinical testing should be performed to determine whether expanded CD133(+) cells have any clinical advantages over purified CD133(+) cells.
authors.affiliation: Pontifícia Universidade Católica do Paraná, Institute for Health and Biological Sciences, Rua Imaculada Conceição, 1155 Curitiba, Paraná, 80215901, Brazil. alexandra.senegaglia@pucpr.br
authors.forename: Alexandra C
authors.initials: AC
authors.lastname: Senegaglia
authors.forename: Laura A
authors.initials: LA
authors.lastname: Barboza
authors.forename: Bruno
authors.initials: B
authors.lastname: Dallagiovanna
authors.forename: Carlos A M
authors.initials: CA
authors.lastname: Aita
authors.forename: Paula
authors.initials: P
authors.lastname: Hansen
authors.forename: Carmen L K
authors.initials: CL
authors.lastname: Rebelatto
authors.forename: Alessandra M
authors.initials: AM
authors.lastname: Aguiar
authors.forename: Nelson I
authors.initials: NI
authors.lastname: Miyague
authors.forename: Patrícia
authors.initials: P
authors.lastname: Shigunov
authors.forename: Fabiane
authors.initials: F
authors.lastname: Barchiki
authors.forename: Alejandro
authors.initials: A
authors.lastname: Correa
authors.forename: Marcia
authors.initials: M
authors.lastname: Olandoski
authors.forename: Marco A
authors.initials: MA
authors.lastname: Krieger
authors.forename: Paulo R S
authors.initials: PR
authors.lastname: Brofman
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001483:Base Sequence; D002196:Capillaries / Q000254:growth & development; D002454:Cell Differentiation; D049109:Cell Proliferation; D017931:DNA Primers / Q000235:genetics; D042783:Endothelial Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D018189:Immunomagnetic Separation; D066298:In Vitro Techniques; D007231:Infant, Newborn; D008297:Male; D009203:Myocardial Infarction / Q000503:physiopathology / Q000628:therapy*; D018919:Neovascularization, Physiologic; D010455:Peptides / Q000378:metabolism*; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D033581:Stem Cell Transplantation; D014183:Transplantation, Heterologous; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D016277:Ventricular Function, Left
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D017931:DNA Primers; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606025:Prom1 protein, rat; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A