白膜法制备手工血小板方法的探讨

目的通过研究采集与制备时间对手工血小板质量的影响,探讨白膜法制备手工血小板的方法。方法采集28例健康献血者400ml全血,采用（两步法）白膜法分离制备手工血小板,按照采集与制备时间的不同均分为4组,每组7例。对照组：新鲜全血采集6 h之内立即进行手工血小板的分离制备（对照组）;B组：新鲜全血采集6 h之内进行第一步分离,白膜放置6 h进行第二步分离;C组：新鲜全血采集6 h之内进行第一步分离,白膜放置24 h进行第二步分离;D组：新鲜全血采集6～8 h之后进行手工血小板的分离制备。检测血小板计数（platelet count,PLT）、血小板平均体积（mean platelet volum,MPV）、白细胞残余率、pH值、低渗休克反应（hypotonic shock response,HSR）、血小板聚集率以及血小板膜P-选择素（CD62p）的变化。结果白膜放置6 h之后（B组）,制备手工血小板其血小板计数（779.9±83.8）×109/L和HSR（62.61±5.24）%高于其他3组;MPV（6.44±0.17）fl和CD62p（47.67±7.40）%低于其他3组。B、C、D3组血小板计数、pH值、HSR和血小板聚集的比较均优于对照组,且差异有统计学意义（P〈0.05）,4组CD62p流式细胞术检测结果无统计学差异。结论手工制备血小板可以打破传统6～8 h之内必须制备的概念,尤以新鲜全血立即制备第一步,白膜放置6 h之后制备手工血小板为最优。

Exploration on Preparation Method of Buffy Coat-platelet Concentrates

Objective To study the effect of collection time and preparation time on the quality of buffy coat-platelet concentrates(BC-PC) and explore the preparation method of BC-PC. Methods 400 ml of whole blood were collected from 28 healthy donors,and then were separated and prepared to obtain the platelet though two-step buffy coat method.The samples were divided into four groups(7 samples in each group) according to the collection time and preparation time.In group A(control),platelet concentrates(PCs) were prepared from fresh whole blood within 6 hours on the day of collection.In group B,PCs were prepared from fresh whole blood within 6 hours,subsequently stored for 6 hours,and then underwent the second step of separation.In group C,PCs were prepared from fresh whole blood within 6hours,stored for 24 hours,and then underwent the second step of separation.In group D,PCs were prepared from the whole blood between 6 to 8 hours.Platelet count(PLT),mean platelet volume(MPV),white blood cell residual rate,pH value,hypotonic shock response(HSR),platelet aggregation and the CD62p expression were detected. Results The group B had the highest PLT count((779.9±83.8)×109/L)) and HSR((62.61±5.24)%) and lowest MPV((6.44±0.17)fl) and CD62P expression.The platelet count,pH value,HSR and platelet aggregation of groups B,C and D were significantly better than that of group A(P<0.05).There was no difference among the four groups in the CD62p expression detected by flow cytometry. Conclusion PCs can be produced from the whole blood by the BC method after being stored for 6 to 8 hours.PCs are best prepared by BC method with fresh whole blood preparation as the first step and PLT preparation after 6 hours of buffy coat storing.