Néron Sonia, Thibault Louis, Dussault Nathalie, Côté Geneviève, Ducas Eric, Pineault Nicolas, Roy Annie
Héma-Québec, Research and Development, Québec, Quebec, Canada.
Transfusion. 2007 Jun;47(6):1042-9. doi: 10.1111/j.1537-2995.2007.01233.x.
The yield of white blood cells (WBCs) extracted from whole-blood leukoreduction filters can be affected by the storage conditions and delay before filtration. Platelets (PLTs) collected with apheresis instruments (Trima Accel, Gambro BCT) are leukoreduced during the procedure on a fluidized particle bed in a leukoreduction chamber (LRS chamber). In this report, the residual cell content of these LRS chambers was characterized to determine whether it would be a valuable source of viable human blood cells.
The content of LRS chambers was eluted by gravity, and peripheral blood mononuclear cells (PBMNCs) were purified on a Ficoll-Paque gradient. Analyses were performed before and after freezing. Proportions of CD3+, CD14+, CD16+, CD19+, CD34+, and CD45+ cells were determined by flow cytometry. The frequency of T cells expressing CD4, CD8, and CD27 and of B cells expressing immunoglobulin G (IgG), IgM, and CD27 was also determined.
LRS chambers held approximately 10(9) CD45+ cells representing the normal proportions of CD3+, CD14+, CD16+, and CD19+ cell populations of PBMNCs. A small fraction of these CD45+ cells were CD34+CD38+ cells (0.3 +/- 0.2%). The viability of these cells, measured before and after freezing, was more than 95 percent.
The residual cell content of Trima Accel LRS chambers recovered after PLT collection is a good source of viable monocytes and lymphocytes. These PBMNCs, containing CD3+, CD14+, CD16+, CD19+, and CD34+ cells can be frozen to prepare cell banks, which opens new avenues for utilization in several physiologic studies or even in cellular therapy applications.
从全血白细胞滤除器中提取的白细胞(WBC)产量可能会受到储存条件和过滤前延迟时间的影响。使用血液成分分离仪器(Trima Accel,甘布罗BCT公司)采集的血小板(PLT)在程序中于白细胞滤除室(LRS室)的流化颗粒床上进行白细胞滤除。在本报告中,对这些LRS室的残留细胞成分进行了表征,以确定其是否会成为有活力的人类血细胞的宝贵来源。
通过重力洗脱LRS室的内容物,并在Ficoll-Paque梯度上纯化外周血单核细胞(PBMNC)。在冷冻前后进行分析。通过流式细胞术测定CD3 +、CD14 +、CD16 +、CD19 +、CD34 +和CD45 +细胞的比例。还测定了表达CD4、CD8和CD27的T细胞以及表达免疫球蛋白G(IgG)、IgM和CD27的B细胞的频率。
LRS室含有约10⁹个CD45 +细胞,代表PBMNC中CD3 +、CD14 +、CD16 +和CD19 +细胞群体的正常比例。这些CD45 +细胞中的一小部分是CD34 + CD38 +细胞(0.3±0.2%)。这些细胞在冷冻前后测量的活力超过95%。
血小板采集后回收的Trima Accel LRS室的残留细胞成分是有活力的单核细胞和淋巴细胞的良好来源。这些含有CD3 +、CD14 +、CD16 +、CD19 +和CD34 +细胞的PBMNC可以冷冻以制备细胞库,这为在多项生理学研究甚至细胞治疗应用中的利用开辟了新途径。