Vasculogenic conditioning of peripheral blood mononuclear cells promotes endothelial progenitor cell expansion and phenotype transition of anti-inflammatory macrophage and T lymphocyte to cells with regenerative potential.
Vasculogenic conditioning of peripheral blood mononuclear cells promotes endothelial progenitor cell expansion and phenotype transition of anti-inflammatory macrophage and T lymphocyte to cells with regenerative potential.
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DOI：
10.1161/jaha.113.000743
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发表时间：
2014-06-25
影响因子：
5.4
通讯作者：
Asahara T
中科院分区：
文献类型：
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作者：
Masuda H;Tanaka R;Fujimura S;Ishikawa M;Akimaru H;Shizuno T;Sato A;Okada Y;Iida Y;Itoh J;Itoh Y;Kamiguchi H;Kawamoto A;Asahara T
涉及单核细胞（MNC）的基于细胞的疗法已被开发用于血管再生以治疗缺血性疾病；然而，治疗性跨国公司的质量控制尚未得到评估。我们研究了外周血 (PB) 跨国公司的治疗潜力，这些跨国公司通过最近开发的内皮祖细胞 (EPC) 定性和数量 (QQ) 培养物进行操作。 PBs是从健康志愿者身上收集的；从这些 PB 中分离的外周血单核细胞 (PBMNC) 用含有干细胞因子、血小板生成素、Flt-3 配体、血管内皮生长因子和白介素-6 的培养基进行 QQ 培养 7 天。在 EPC 集落形成测定中，所得细胞 (QQMNC) 产生的 EPC 集落明显比 PBMNC 更明确。在流式细胞术中，QQMNC 的巨噬细胞和辅助 T 淋巴细胞在表型上极化为血管生成、抗炎和再生亚群：经典 M1 到替代 M2； T 辅助细胞 (Th)1 至 Th2；血管生成或调节性 T 细胞扩增。定量实时聚合酶链反应（qRT-PCR）分析揭示了 QQMNC 与 PBMNC 中主要的促血管生成基因表达。使用小鼠缺血后肢模型，将 QQMNC 肌内移植 (Tx) 的疗效与 PBMNCTx、培养的“早期 EPC”Tx (eEPCTx) 和粒细胞集落刺激因子动员的 CD34+ 细胞 Tx (GmCD34Tx) 进行比较。激光多普勒成像显示，QQMNCTx 后缺血后肢的血液灌注恢复优于 PBMNCTx 和 eEPCTx 后，但也早于 GmCD34Tx 后。缺血后肢的组织学评估和 qRT-PCR 检测表明，与 GmCD34Tx 类似，QQMNCTx 可以增强血管生成和肌肉生成，而与 PBMNCTx 和 eEPCTx 相比，它更能抑制炎症和纤维化。 QQ培养增强PBMCs促进损伤组织再生的能力；考虑到可行的细胞制备，QQ 培养处理的 PBMNC 可能为缺血性疾病提供有前景的治疗选择。网址：irb.med.u-tokai.ac.jp/d/2/monthly/2010.html； IRB 编号：10R-020。网址：irb.med.u-tokai.ac.jp/d/2/monthly/201312.html； IRB 编号：13R228。
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). 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. 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. 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.