Mastronardi R, Cleophax S, Begué S, Hurtado-Nedelec M, Gross S, Bocquet T, Djoudi R
Établissement français du sang- Île-de-France, avenue de l'Île-de-France, 95300 Pontoise, France.
Établissement français du sang, 20, avenue du Stade-de-France, 93218 Saint-Denis, France.
Transfus Clin Biol. 2019 Sep;26(3):164-170. doi: 10.1016/j.tracli.2019.06.188. Epub 2019 Jun 18.
The collection of granulocytes by apheresis requires volunteer donor stimulation by corticoids and the use of HES, a compound which is currently challenged by potential safety issues. Preparation of pooled granulocytes concentrates from whole blood buffy coats (PGC) represent an alternative to apheresis with a better benefit/risk for the donors.
Whole blood is collected in a bottom and top blood bag for buffy coat preparation. After centrifugation and separation, buffy coat are obtained. Twenty ABO matched buffy coats are selected for processing into one PGC. Four pools of five buffy coats were made, platelet additive solution is added to each pool, mixed gently and centrifuged. The red cell residue, supernatant and granulocyte rich layer are separated. Two granulocyte rich layers are pooled and added with 70mL of ABO matched plasma from the initial donations (=PGC10). The final PGC (=PGC20) is obtained by pooling two PGC10 into a platelet storage bag. Neutrophil content and in-vitro functionality are assessed at day of preparation (D1) and at expiry hour, 48 hours after collection (D2).
On N=18, mean: Volume=408±4mL, 2.21010±0.24 neutrophils, Hematocrit=18%±3%, 4.71011platelets. Viability is well preserved: 95%±6% day of PGC preparation, 85%±7% after 24h of storage (D2). Functionality (ROS production measurement) is well preserved: 1.36±0.25 at D1 and 1.38±0.18 at D2. Expression and modulation of adhesion molecules after stimulation are normal at D1 and slightly decreased at D2 but still normal.
PGC20 in vitro characteristics are in conformance with the EDQM guide (V19) and similar to apheresis for granulocytes content and hematocrit. The viability and two mean indicators which explore neutrophil function are well maintained during PGC preparation and after 24 hours of storage.
通过单采术采集粒细胞需要用皮质类固醇刺激志愿捐献者，并使用羟乙基淀粉（HES），而该化合物目前面临潜在的安全问题。从全血白膜层制备混合粒细胞浓缩物（PGC）是单采术的一种替代方法，对捐献者而言其风险效益比更佳。
将全血采集到用于制备白膜层的上下两层血袋中。经过离心和分离后，获得白膜层。选择20份ABO血型匹配的白膜层用于制备一份PGC。将五份白膜层制成四组，向每组中加入血小板添加剂溶液，轻轻混合并离心。分离出红细胞残余物、上清液和富含粒细胞的层。将两层富含粒细胞的层合并，并加入70mL来自初始捐献的ABO血型匹配血浆（=PGC10）。通过将两份PGC10合并到一个血小板储存袋中获得最终的PGC（=PGC20）。在制备当天（D1）和采集后48小时的有效期（D2）评估中性粒细胞含量和体外功能。
在N = 18时，平均值：体积 = 408±4mL，中性粒细胞2.2×10¹⁰±0.24，血细胞比容 = 18%±3%，血小板4.7×10¹¹。活力保存良好：PGC制备当天为95%±6%，储存24小时后（D2）为85%±7%。功能（活性氧产生测量）保存良好：D1时为1.36±0.25，D2时为1.38±0.18。刺激后黏附分子的表达和调节在D1时正常，在D2时略有下降但仍正常。
PGC20的体外特性符合欧洲药品质量管理局指南（V19），在粒细胞含量和血细胞比容方面与单采术相似。在PGC制备过程中和储存24小时后，活力以及两个探索中性粒细胞功能的平均指标均得到良好维持。
