RhG-CSF Mobilized and Apheresis-Collected Endothelial Progenitor Cells for Therapeutic Vasculogenesis.
RhG-CSF Mobilized and Apheresis-Collected Endothelial Progenitor Cells for Therapeutic Vasculogenesis.
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用于治疗性血管生成的 RhG-CSF 动员和血浆分离术收集的内皮祖细胞。
DOI：
10.1182/blood.v106.11.298.298
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Z. Estrov
中科院分区：
文献类型：
--
作者：
M. Korbling;J. Reuben;Hui Gao;Bang‐Ning Lee;S. Giralt;I. Khouri;R. Saliba;R. Champlin;Z. Estrov
内皮祖细胞 (EPC) 已被鉴定为造血组织来源的祖细胞的一部分，由于组织损伤而被动员到外周血 (PB) 中。因此，循环中的 EPC 可能通过帮助缺血组织的新血管形成而具有治疗潜力。本研究的目的是提供关于稳定状态下和rhG-CSF动员后循环EPC的可用性以及通过白细胞分离术收集它们的临床数据。八名健康捐赠者接受了为期 4 天的 rhG-CSF (10 μg/kg) 治疗。在 rhG-CSF 治疗的第四天进行连续流动白细胞去除术（COBE Spectra，版本 4.7）。在开始rhG-CSF治疗之前、在血浆分离术之前以及从血浆分离术收集袋中抽取血样。每次单采术收集的总细胞数基于处理中位数 17 L [10.7–20.1] 血量，或大约是捐献者总血量的三倍。通过流式细胞术分析血液和单采血液样本中的 EPC 表面标志物 CD34、CD133、VEGFR-2，并通过形成 EPC 集落 (CFU-EPC) 进行分析。 Wilcoxon 配对符号秩检验用于比较不同采样点的分布。所有 CD133 亚群均为 CD14 阴性，以排除分化的单核细胞或巨噬细胞。循环 CD34 + CD133 + VEGFR-2 + 细胞的中位稳态 PB 浓度为 0.9/μl [0–3]，CD34 + 133 – VEGFR-2 + EPC 为 1.7/μl [0.9–4]。 rhG-CSF 动员治疗 4 天后，PB CD34 + CD133 + VEGFR-2 + 子集浓度中位数增加了 8 倍 [p < 0.01]，CD34 + 133 – VEGFR-2 + 子集浓度中位数较基线增加了 10 倍 [p < 0.01]。循环 CFU-EPC 的中位 PB 浓度增加了 10 倍 [p < 0.02]。每公斤体重通过白细胞去除术收集的 CD34 + CD133 + VEGFR-2 + 和 CD34 + CD133 - VEGFR-2 + 细胞的中位绝对数分别为 0.845 x 10 6 [0.12–2.55] 和 1.54 x 10 6 [0.06–2.4]。在稳态和动员的 PB 以及单采血液成分中鉴定出一小群 CD133 + 34 − VEGFR-2 + 细胞 (0.11 x 10 6 /kg [0–0.26])。 CD34 - 表达 CD133 的祖细胞被认为代表含有 SCID 再生细胞的原始祖细胞群。这些未成熟细胞的 VEGFR-2 共表达可能定义了有助于血管生成的循环细胞群。我们的数据表明，克隆形成的循环 EPC 可以通过 rhG-CSF 动员，并通过连续流白细胞分离术收集，产生大量 EPC，特别是对于 70 kg 的个体，每次白细胞分离术可产生 60 x 10 6 EPCs。循环 EPC 代表了一种新型血细胞成分，可在临床上大量使用（未经操作或经过 EPC 选择）用于治疗性血管生成。
Endothelial progenitor cells (EPCs) have been identified as part of hematopoietic tissue-derived progenitor cells that are mobilized into the peripheral blood (PB) as a result of tissue injury. It therefore seems likely that circulating EPCs have therapeutic potential by aiding in the neovascularization of ischemic tissue. It was the purpose of this study to provide clinical data on the availability of circulating EPCs at steady-state and after rhG-CSF mobilization, and their collection by leukapheresis. Eight healthy donors underwent rhG-CSF (10 μg/kg) treatment over 4 days. Continuous-flow leukapheresis (COBE Spectra, Version 4.7) was performed on the fourth day of rhG-CSF treatment. Blood samples were drawn before starting rhG-CSF treatment, before apheresis, and from the apheresis collection bag. The total cell numbers collected per apheresis were based on processing a median of 17 L [10.7–20.1] blood volume or approximately three times the donor’s total blood volume. Blood and apheresis samples were analyzed by flow cytometry for EPC surface markers CD34, CD133, VEGFR-2, and by forming EPC colonies (CFU-EPC). The Wilcoxon matched-pairs signed-ranks test was used to compare the distributions at various sampling points. All CD133 subsets were CD14 negative to exclude differentiated monocytes or macrophages. The median steady-state PB concentrations of circulating CD34 + CD133 + VEGFR-2 + cells was 0.9/μl [0–3], or 1.7/μl [0.9–4] for CD34 + 133 − VEGFR-2 + EPCs. After 4 days of rhG-CSF mobilization treatment the PB CD34 + CD133 + VEGFR-2 + subset concentration increased by a median of 8-fold [p 0.01], and the CD34 + 133 − VEGFR-2 + subset concentration by a median of 10-fold [p 0.01] over baseline. The median PB concentration of circulating CFU-EPCs increased by 10-fold [p 0.02]. The median absolute number of CD34 + CD133 + VEGFR-2 + and CD34 + CD133 - VEGFR-2 + cells collected by leukapheresis per kg of body weight was 0.845 x 10 6 [0.12–2.55] and 1.54 x 10 6 [0.06–2.4], respectively. A small population of CD133 + 34 − VEGFR-2 + cells was identified in steady state and mobilized PB, and in the apheresis collect (0.11 x 10 6 /kg [0–0.26]). CD34 − progenitor cells expressing CD133 are believed to represent a primitive progenitor cell population containing SCID-repopulating cells. VEGFR-2 coexpression of those immature cells may define a circulating cell population that contributes to vasculogenesis. Our data suggest that clonogenic circulating EPCs can be mobilized by rhG-CSF and collected by continuous-flow leukapheresis generating large numbers of EPCs, specifically in the range of 60 x 10 6 EPCs per leukapheresis procedure for a 70 kg individual. Circulating EPCs represent a novel blood cell component that can be clinically used in large quantities, either unmanipulated or EPC-selected, for therapeutic vasculogenesis.