Investigating the feasibility of stem cell enrichment mediated by immobilized selectins
Investigating the feasibility of stem cell enrichment mediated by immobilized selectins
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DOI：
10.1021/bp0702222
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发表时间：
2007-11-01
影响因子：
2.9
通讯作者：
King, Michael R.
中科院分区：
文献类型：
--
作者：
Charles, Nichola;Liesveld, Jane L.;King, Michael R.
造血干细胞疗法用于治疗恶性和非恶性疾病，并且造血干细胞和祖细胞（HSPC）的富集具有降低移植物抗宿主疾病或复发的可能性的潜力，所述移植物抗宿主疾病或复发是与疗法相关的潜在致命并发症。目前的商业HSPC分离技术仅依赖于CD 34表面标记物，虽然它们已被证明是非常宝贵的，但它们可能非常耗时，并且报告了可变的回收率。我们建议选择素介导的富集可以被证明是一种快速有效的方法，用于从成人骨髓（ABM）中回收HSPCs，其基于滚动速度的差异并且独立于CD 34表达。纯化的CD 34 + ABM细胞和CD 34- ABM细胞在生理壁剪切应力下灌注在固定的P-、E-和L-选择素-IgG上，并收集滚动速度和细胞保留数据。在所有三种选择素上，CD 34 + ABM细胞通常比CD 34- ABM细胞表现出更低的滚动速度和更高的保留。对于初始CD 34 + ABM细胞浓度范围为1%至5%，我们预测纯度增加范围为5.2%至36.1%，这取决于所使用的选择素。此外，选择素介导的细胞富集不限于滚动速度具有固有差异的细胞亚群。CD 34 + KG 1a细胞和CD 34-HL 60细胞在研究的整个剪切应力范围内对固定的P-选择素-IgG表现出几乎相同的滚动速度。然而，当抗CD 34抗体与P-选择素-IgG共固定时，CD 34 + KG 1a细胞的滚动速度显著降低，使得选择素介导的细胞富集成为可行的选择。最佳的细胞富集在固定的选择素表面可以在10分钟内实现，比目前大多数市售系统快得多。
Hematopoietic stem cell therapy is used to treat both malignant and non-malignant diseases, and enrichment of the hematopoietic stem and progenitor cells (HSPCs) has the potential to reduce the likelihood of graft vs host disease or relapse, potentially fatal complications associated with the therapy. Current commercial HSPC isolation technologies rely solely on the CD34 surface marker, and while they have proven to be invaluable, they can be time-consuming with variable recoveries reported. We propose that selectin-mediated enrichment could prove to be a quick and effective method for recovering HSPCs from adult bone marrow (ABM) on the basis of differences in rolling velocities and independently of CD34 expression. Purified CD34+ ABM cells and the unselected CD34- ABM cells were perfused over immobilized P-, E-, and L-selectin-IgG at physiologic wall shear stresses, and rolling velocities and cell retention data were collected. CD34+ ABM cells generally exhibited lower rolling velocities and higher retention than the unselected CD34- ABM cells on all three selectins. For initial CD34+ ABM cell concentrations ranging from 1% to 5%, we predict an increase in purity ranging from 5.2% to 36.1%, depending on the selectin used. Additionally, selectin-mediated cell enrichment is not limited to subsets of cells with inherent differences in rolling velocities. CD34+ KG1a cells and CD34- HL60 cells exhibited nearly identical rolling velocities on immobilized P-selectin-IgG over the entire range of shear stresses studied. However, when anti-CD34 antibody was co-immobilized with the P-selectin-IgG, the rolling velocity of the CD34+ KG1a cells was significantly reduced, making selectin-mediated cell enrichment a feasible option. Optimal cell enrichment in immobilized selectin surfaces can be achieved within 10 min, much faster than most current commercially available systems.