Journal
JOURNAL OF THE AMERICAN HEART ASSOCIATION
Volume 3, Issue 3, Pages -Publisher
WILEY
DOI: 10.1161/JAHA.113.000743
Keywords
anti-inflammation; cell-based therapy; peripheral blood mononuclear cells; serum-free culture; vascular regeneration
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Funding
- Riken Center for Developmental Biology Collaborative Research Fund, Kobe, Japan [08001475]
- Ministry of Health, Labor and Welfare, Japan [25280401]
- Ministry of Education, Culture, Sports, Science and Technology (Academic Frontier Promotion Program), Japan
- Ministry of Education, Culture, Sports, Science and Technology, Japan [25461091]
- Tokai University School of Medicine Research Aid
- Research and Study Project of Tokai University Education System General Research Organization, Japan
- Grants-in-Aid for Scientific Research [26461320, 25461091] Funding Source: KAKEN
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Background-Cell-based therapies involving mononuclear cells (MNCs) have been developed for vascular regeneration to treat ischemic diseases; however, quality control of therapeutic MNCs has not been evaluated. We investigated the therapeutic potential of peripheral blood (PB) MNCs, operated by recently developed quality and quantity (QQ) culture of endothelial progenitor cells (EPCs). Methods and Results-PBs were collected from healthy volunteers; peripheral blood mononuclear cells (PBMNCs) isolated from these PBs were subjected to QQ culture for 7 days with medium containing stem cell factor, thrombopoietin, Flt-3 ligand, vascular endothelial growth factor, and interleukin-6. The resulting cells (QQMNCs) in EPC colony-forming assay generated significantly more definitive EPC colonies than PBMNCs. In flow cytometry, macrophages and helper T lymphocytes of QQMNCs became phenotypically polarized into angiogenic, anti-inflammatory, and regenerative subsets: classical M1 to alternative M2; T helper (Th) 1 to Th2; angiogenic or regulatory T-cell expansion. Quantitative real-time polymerase chain reaction (qRT-PCR) assay revealed the predominant proangiogenic gene expressions in QQMNCs versus PBMNCs. Using murine ischemic hindlimb models, the efficacy of QQMNC intramuscular transplantation (Tx) was compared to that of PBMNCTx, cultured early EPC Tx (eEPCTx), and granulocyte colony-stimulating factor mobilized CD34(+) cell Tx (GmCD34Tx). Laser Doppler imaging revealed the blood perfusion recovery in ischemic hindlimbs after QQMNCTx superior to after PBMNCTx and eEPCTx, but also earlier than after GmCD34Tx. Histological evaluations and qRT-PCR assays in ischemic hindlimbs demonstrated that QQMNCTx, similarly to GmCD34Tx, enhanced angiovasculogenesis and myogenesis, whereas it preponderantly inhibited inflammation and fibrosis versus PBMNCTx and eEPCTx. Conclusions-QQ culture potentiates the ability of PBMNCs to promote regeneration of injured tissue; considering the feasible cell preparation, QQ culture-treated PBMNCs may provide a promising therapeutic option for ischemic diseases.
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