title: Development of serum-free quality and quantity control culture of colony-forming endothelial progenitor cell for vasculogenesis.
abstract: Quantitative and qualitative impairment of endothelial progenitor cells (EPCs) limits the efficacy of autologous cell therapy in patients with cardiovascular diseases. Here, we developed a serum-free quality and quantity control culture system for colony-forming EPCs to enhance their regenerative potential. A culture with serum-free medium containing stem cell factor, thrombopoietin, vascular endothelial growth factor, interleukin-6, and Flt-3 ligand was determined as optimal quality and quantity culture (QQc) in terms of the most vasculogenic colony-forming EPC expansion, evaluated by the newly established EPC colony formation assay. The QQc of umbilical cord blood-CD133(+) cells for 7 days produced a 52.9-fold increase in total cell number and 3.28-fold frequency in definitive EPC colony development, resulting in a 203.9-fold increase in estimated total definitive EPC colony number in vitro. Pre- or post-QQc cells were intramyocardially transplanted into nude rats with myocardial infarction (MI). Echocardiographic and micromanometer-tipped conductance catheter examinations 28 days post-MI revealed significant preservation of left ventricular (LV) function in rats receiving pre- or post-QQc cells compared with those receiving phosphate-buffered saline. Assessments of global LV contractility indicated a dose-dependent effect of pre- or post-QQc cells and the superior potency of post-QQc cells over pre-QQc cells. Furthermore, immunohistochemistry showed more abundant formation of both human and rat endothelial cells and cardiomyocytes in the infarcted myocardium following transplantation of post-QQc cells compared with pre-QQc cells. Our optimal serum-free quality and quantity culture may enhance the therapeutic potential of EPCs in both quantitative and qualitative aspects for cardiovascular regeneration.
authors.affiliation: Department of Regenerative Medicine Science, Tokai University School of Medicine, Isehara, Japan.
authors.forename: Haruchika
authors.initials: H
authors.lastname: Masuda
authors.forename: Hiroto
authors.initials: H
authors.lastname: Iwasaki
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Kawamoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akimaru
authors.forename: Masakazu
authors.initials: M
authors.lastname: Ishikawa
authors.forename: Masaaki
authors.initials: M
authors.lastname: Ii
authors.forename: Tomoko
authors.initials: T
authors.lastname: Shizuno
authors.forename: Atsuko
authors.initials: A
authors.lastname: Sato
authors.forename: Rie
authors.initials: R
authors.lastname: Ito
authors.forename: Miki
authors.initials: M
authors.lastname: Horii
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Ishida
authors.forename: Shunichi
authors.initials: S
authors.lastname: Kato
authors.forename: Takayuki
authors.initials: T
authors.lastname: Asahara
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002021:Buffers; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods* / Q000592:standards; D049109:Cell Proliferation; D064987:Cell- and Tissue-Based Therapy / Q000379:methods / Q000592:standards; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay / Q000379:methods* / Q000592:standards; D016895:Culture Media, Serum-Free / Q000378:metabolism*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004452:Echocardiography; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D009200:Myocardial Contraction; D009203:Myocardial Infarction / Q000378:metabolism / Q000628:therapy; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D018919:Neovascularization, Physiologic; D010455:Peptides / Q000378:metabolism; D011786:Quality Control; D051381:Rats; D011923:Rats, Nude; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D016277:Ventricular Function, Left
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D002021:Buffers; D016895:Culture Media, Serum-Free; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606025:Prom1 protein, rat