ABSTRACT
【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.