In:
Journal of the American Heart Association, Ovid Technologies (Wolters Kluwer Health), Vol. 3, No. 3 ( 2014-05-22)
Abstract:
Cell‐based therapies involving mononuclear cells ( MNC s) have been developed for vascular regeneration to treat ischemic diseases; however, quality control of therapeutic MNC s has not been evaluated. We investigated the therapeutic potential of peripheral blood ( PB ) MNC s, operated by recently developed quality and quantity ( QQ ) culture of endothelial progenitor cells ( EPC s). Methods and Results PBs were collected from healthy volunteers; peripheral blood mononuclear cells ( PBMNC s) isolated from these PB s 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 ( QQMNC s) in EPC colony‐forming assay generated significantly more definitive EPC colonies than PBMNC s. In flow cytometry, macrophages and helper T lymphocytes of QQMNC s 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 QQMNC s versus PBMNC s. 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 eEPCT x, but also earlier than after GmCD34Tx. Histological evaluations and q RT ‐ 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 PBMNC s to promote regeneration of injured tissue; considering the feasible cell preparation, QQ culture‐treated PBMNC s may provide a promising therapeutic option for ischemic diseases. Clinical Trial Registration URL : irb.med.u-tokai.ac.jp/d/2/monthly/2010.html ; IRB No.: 10R‐020. URL : irb.med.u-tokai.ac.jp/d/2/monthly/201312.html ; IRB No.: 13R228.
Type of Medium:
Online Resource
ISSN:
2047-9980
DOI:
10.1161/JAHA.113.000743
Language:
English
Publisher:
Ovid Technologies (Wolters Kluwer Health)
Publication Date:
2014
detail.hit.zdb_id:
2653953-6
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