改良顶底法在脐带血有核细胞分离中的优化

Optimization of the new Plates-B&T method for nucleated cell separation in cord blood

目的 探索使用改良顶底法分离脐带血有核细胞,减少分离过程中白膜层的位移扰动,提高分离效果。方法 在顶底法的基础上,采用两块透明板材,中间使用螺丝连接,离心时将血袋夹在中间,离心完成后带装置整体取出放入分浆夹中进行分离,优化分离参数,形成改良顶底法,并与传统富浆法进行平行比较。结果 改良顶底法使用夹板可以有效避免离心后白膜层位移,较好地解决了液面扰动问题。改良顶底法只需一次离心,与富浆法比较,离心分离时间由(29.12±2.79)min降至(14.91±2.12)min(P<0.05)。改良顶底法的TNC回收率为(89.04±5.19)%,虽较富浆法有小幅提升,但无统计学差异(P>0.05)。改良顶底法RBC去除率为(55.56±22.57)%,HCT为(0.48±0.09)L/L,较富浆法均有统计学差异(P<0.05),在中、低体积段中的差异更大。结论 改良顶底法对顶底法进行了优化,提高了脐带血分离效率和质量。

Objective To optimize the bottom and top(B&T) method for nucleated cell separation in cord blood in order to reduce the disturbance and improve the nucleated cell yield.Methods A new device comprised of two plastic plates and connection units was developed.B&T Cord blood bag was put between the plates,which were then centrifuged and put into the serum expresser together to go through serum and red blood cells separation simultaneously.After centrifugation parameter adjustment,the new Plates-B&T method was compared with traditional method in parallel.Results New Plates-B&T method could avoid buffy coat movement and disturbance.Compared to conventional PRP method,Plates-B&T method reduced centrifugation and separation time from(29.12±2.79)min to(14.91±2.12)min(P<0.05).Using new method,the recovery rate of total nucleated cell(TNC) was(89.04±5.19)%,the removal rate of red blood cell(RBC) was(55.56±22.57) %,and hematocrit(HCT) in buffy coat was(0.48±0.09)L/L.RBC removal was enhanced significantly,especially for cord blood samples with medium-low volume.Conclusion Based on B&T method,we established a new device and method,which increased the efficiency and quality of cord blood nucleated cell separation.