Tsang K S, Li C K, Wong A P, Leung Y, Lau T T, Li K, Shing M M, Chik K W, Yuen P M
Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, Sha Tin, Hong Kong, People's Republic of China. @
Transfusion. 1999 Nov-Dec;39(11-12):1212-9. doi: 10.1046/j.1537-2995.1999.39111212.x.
Various open and semi-closed methods are used for red cell (RBC) depletion and hematopoietic progenitor cell (HPC) enrichment of bone marrow (BM) in vitro, but with variable efficacy. A simple, efficient, and safe method using dextran 110k was developed.
An equal volume of 4.5-percent dextran was applied to major ABO-incompatible BM in transfer bags and sedimentation was allowed for 30 minutes. RBCs, nucleated cells (NCs), and mononuclear cells (MNCs) from BM allografts before and after dextran sedimentation (DS) were counted. Flow cytometry, short-term cultures, and long-term cultures were performed to assay the respective recovery of CD34+ cells, colony-forming units (CFUs), and long-term culture-initiating cells (LTC-ICs).
Sixteen BM collections were processed. The mean volume was 666 mL (range, 189-1355 mL). The mean +/-1 SD post-DS NC, MNC, CD34+ cell, and CFU counts per kg of the recipient's body weight were 4.11 +/-1.74 x 10(8), 8.98 +/- 3.68 x 10(7), 2.90 +/- 1.95 x 10(6), and 2.03 +/- 2.01 x 10(5), respectively, with the corresponding post-DS recovery being 90.6 percent, 90 percent, 92.4 percent, and 100.8 percent. The numbers of LTC-ICs in cultures (up to 12 weeks) of pre-DS and post-DS samples of five BM allografts were comparable (p = 0.91). Residual RBCs were 5.1 +/- 4.6 (0.1-14) mL with depletion of 96.5 +/- 3.2 percent. There was no significant difference in the mean absolute RBC count in post-DS BM allografts and in four ficoll-treated BM allografts (8.09 x 10(10) vs. 4.9 x 10(9); p = 0.206) and in eight major ABO-incompatible peripheral blood HPC collections (8.09 x 10(10) vs. 9.81 x 10(10); p = 0.87). No posttransplant hemolysis was encountered. Engraftment occurred at 22 +/- 7 days, which is similar to that of four transplants with ficoll-treated BM allografts (22 +/- 9; p = 0.611) and 54 unprocessed BM allografts (19 +/- 6; p = 0.129).
DS is an efficient method of depleting RBCs in major ABO-incompatible BM allografts without significant loss of HPCs.
体外进行红细胞（RBC）去除及骨髓（BM）造血祖细胞（HPC）富集时，使用了多种开放式和半封闭式方法，但效果各异。研发出一种使用110k右旋糖酐的简单、高效且安全的方法。
将等体积的4.5%右旋糖酐应用于转移袋中主要ABO血型不相合的骨髓，静置沉降30分钟。对右旋糖酐沉降（DS）前后骨髓移植物中的红细胞、有核细胞（NC）及单核细胞（MNC）进行计数。采用流式细胞术、短期培养及长期培养来检测CD34+细胞、集落形成单位（CFU）及长期培养起始细胞（LTC-IC）各自的回收率。
共处理了16份骨髓采集样本。平均体积为666 mL（范围189 - 1355 mL）。DS后每千克受者体重的NC、MNC、CD34+细胞及CFU计数的均值±1标准差分别为4.11±1.74×10⁸、8.98±3.68×10⁷、2.90±1.95×10⁶及2.03±2.01×10⁵，相应的DS后回收率分别为90.6%、90%、92.4%及100.8%。五份骨髓移植物DS前和DS后样本培养（长达12周）中的LTC-IC数量相当（p = 0.91）。残余红细胞为5.1±4.6（0.1 - 14）mL，去除率为96.5±3.2%。DS后骨髓移植物与四份经聚蔗糖处理的骨髓移植物中的平均绝对红细胞计数无显著差异（8.09×10¹⁰对4.9×10⁹；p = 0.206），与八份主要ABO血型不相合的外周血HPC采集样本中的也无显著差异（8.09×10¹⁰对9.81×10¹⁰；p = 0.87）。未发生移植后溶血。植入发生于22±7天，这与四份经聚蔗糖处理的骨髓移植物移植情况（22±9；p = 0.611）及54份未处理的骨髓移植物移植情况（19±6；p = 0.129）相似。
DS是去除主要ABO血型不相合骨髓移植物中红细胞的有效方法，且造血祖细胞无显著损失。
