Efficient removal of platelets from peripheral blood progenitor cell products using a novel micro-chip based acoustophoretic platform.
Efficient removal of platelets from peripheral blood progenitor cell products using a novel micro-chip based acoustophoretic platform.
复制标题
DOI：
10.1371/journal.pone.0023074
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Scheding S
中科院分区：
文献类型：
--
作者：
Dykes J;Lenshof A;Åstrand-Grundström IB;Laurell T;Scheding S
过度采集血小板是当前基于离心的外周血祖细胞（PBPC）单采中的一种不良副作用。我们研究了一种新的基于微芯片的声电泳技术，利用超声驻波力从PBPC产品中去除血小板。通过将声驻波场施加到微通道中连续流动的细胞悬浮液上，可以根据细胞的物理性质将细胞与周围介质分离。从患者（n = 15）和健康供体（n = 6）获得PBPC样品，并在声发射芯片上分选。 设置声力以将白细胞从血小板分别分离成目标级分和废物级分。分析PBPC样品、目标和废物级分的细胞回收率、纯度和功能性。白细胞与目标级分的中位分离效率为98%，而血小板被有效去除89%。PBPC样品和相应的目标组分在CD 34+造血祖细胞/干细胞百分比以及白细胞/淋巴细胞亚群分布方面相似。在PBPC样品、目标和废物级分中，中位活力分别为98%、98%和97%。来自造血祖细胞测定的结果表明分选后保留的集落形成能力。通过P-选择素（CD 62 P）表达评估血小板活化显示，与PBPC输入样品（9%）相比，靶组分（19%）和废物组分（20%）中的CD 62 P+血小板分别显著增加。然而，与储存的血库血小板浓缩物（48%）相比，活化率较低。声泳可用于有效地消耗PBPC样品中的血小板，同时保留靶干/祖细胞和白细胞群体、细胞活力和祖细胞集落形成能力。因此，声电泳是一种改进当前细胞处理方法的有趣技术。
Excessive collection of platelets is an unwanted side effect in current centrifugation-based peripheral blood progenitor cell (PBPC) apheresis. We investigated a novel microchip-based acoustophoresis technique, utilizing ultrasonic standing wave forces for the removal of platelets from PBPC products. By applying an acoustic standing wave field onto a continuously flowing cell suspension in a micro channel, cells can be separated from the surrounding media depending on their physical properties. PBPC samples were obtained from patients (n = 15) and healthy donors (n = 6) and sorted on an acoustophoresis-chip. The acoustic force was set to separate leukocytes from platelets into a target fraction and a waste fraction, respectively. The PBPC samples, the target and the waste fractions were analysed for cell recovery, purity and functionality. The median separation efficiency of leukocytes to the target fraction was 98% whereas platelets were effectively depleted by 89%. PBPC samples and corresponding target fractions were similar in the percentage of CD34+ hematopoetic progenitor/stem cells as well as leukocyte/lymphocyte subset distributions. Median viability was 98%, 98% and 97% in the PBPC samples, the target and the waste fractions, respectively. Results from hematopoietic progenitor cell assays indicated a preserved colony-forming ability post-sorting. Evaluation of platelet activation by P-selectin (CD62P) expression revealed a significant increase of CD62P+ platelets in the target (19%) and waste fractions (20%), respectively, compared to the PBPC input samples (9%). However, activation was lower when compared to stored blood bank platelet concentrates (48%). Acoustophoresis can be utilized to efficiently deplete PBPC samples of platelets, whilst preserving the target stem/progenitor cell and leukocyte cell populations, cell viability and progenitor cell colony-forming ability. Acoustophoresis is, thus, an interesting technology to improve current cell processing methods.
登录
查看更多内容
影响因子：
2.9
作者：
Hagberg, IA;Akkok, ÇA;Kjeldsen-Kragh, J
通讯作者：
Kjeldsen-Kragh, J
影响因子：
2.7
作者：
Krailadsiri, P;Seghatchian, J
通讯作者：
Seghatchian, J
影响因子：
6.1
作者：
Nilsson, A;Petersson, F;Laurell, T
通讯作者：
Laurell, T
影响因子：
6.1
作者：
Hawkes, JJ;Barber, RW;Coakley, WT
通讯作者：
Coakley, WT
影响因子：
20.3
作者：
Evangelista, V;Manarini, S;Cerletti, C
通讯作者：
Cerletti, C