Wojciechowski Joel C, Narasipura Srinivas D, Charles Nichola, Mickelsen Deanne, Rana Kuldeeepsinh, Blair Martha L, King Michael R
Department of Biomedical Engineering, University of Rochester, Rochester, NY 14627, USA.
Br J Haematol. 2008 Mar;140(6):673-81. doi: 10.1111/j.1365-2141.2007.06967.x. Epub 2008 Jan 22.
Clinical infusion of haematopoietic stem and progenitor cells (HSPCs) is vital for restoration of haematopoietic function in many cancer patients. Previously, we have demonstrated an ability to mimic physiological cell trafficking in order to capture CD34-positive (CD34+) HSPCs using monolayers of the cell adhesion protein P-selectin in flow chambers. The current study aimed to determine if HSPCs could be captured directly from circulating blood in vivo. Vascular shunt prototypes, coated internally with P-selectin, were inserted into the femoral artery of rats. Blood flow through the cell capture device resulted in a wall shear stress of 4-6 dynes/cm(2). After 1-h blood perfusion, immunofluorescence microscopy and flow cytometric analysis revealed successful capture of mononuclear cells positive for the HSPC surface marker CD34. Purity of captured CD34+ cells showed sevenfold enrichment over levels found in whole blood, with an average purity of 28%. Robust cell capture and HSPC enrichment were also demonstrated in devices that were implanted in a closed-loop arterio-venous shunt conformation for 2 h. Adherent cells were viable in culture and able to differentiate into burst-forming units. This study demonstrated an ability to mimic the physiological arrest of HSPCs from blood in an implantable device and may represent a practical alternative for adult stem cell capture and enrichment.
临床输注造血干细胞和祖细胞(HSPCs)对于许多癌症患者恢复造血功能至关重要。此前,我们已证明能够模拟生理细胞运输,以便在流动腔室中使用细胞粘附蛋白P-选择素单层捕获CD34阳性(CD34+)HSPCs。当前研究旨在确定HSPCs是否能够在体内直接从循环血液中捕获。内部涂有P-选择素的血管分流原型被插入大鼠股动脉。通过细胞捕获装置的血流产生了4-6达因/平方厘米的壁面剪应力。在1小时血液灌注后,免疫荧光显微镜和流式细胞术分析显示成功捕获了HSPC表面标志物CD34阳性的单核细胞。捕获的CD34+细胞纯度比全血中发现的水平富集了7倍,平均纯度为28%。在以闭环动静脉分流构型植入2小时的装置中也证明了强大的细胞捕获和HSPC富集。贴壁细胞在培养中具有活力,并能够分化为爆式集落形成单位。这项研究证明了在可植入装置中模拟HSPCs从血液中生理停滞的能力,可能代表了成人干细胞捕获和富集的一种实用替代方法。