PROCESSING AND CRYOPRESERVATION OF PLACENTAL UMBILICAL-CORD BLOOD FOR UNRELATED BONE-MARROW RECONSTITUTION
PROCESSING AND CRYOPRESERVATION OF PLACENTAL UMBILICAL-CORD BLOOD FOR UNRELATED BONE-MARROW RECONSTITUTION
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DOI：
10.1073/pnas.92.22.10119
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发表时间：
1995-10-24
影响因子：
11.1
通讯作者：
STEVENS, CE
中科院分区：
文献类型：
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作者：
RUBINSTEIN, P;DOBRILA, L;STEVENS, CE
胎盘/脐带血（PCB）移植造血恢复的临床证据表明，PCB可以是一个有用的来源，造血干细胞的常规骨髓重建。在不相关的设置，人类白细胞抗原（HLA）匹配的供体必须获得候选患者，因此，冷冻HLA型PCB单位的大面板必须建立，大体积的未经处理的单位，主要由红细胞，血浆，和冷冻保存介质，造成了严重的困难，在这方面的努力，因为在液氮中的存储空间是有限的和昂贵的。我们在这里报告说，几乎所有的造血集落形成细胞存在于PCB单位可以回收在一个统一的体积为20毫升，通过使用rouleaux形成诱导羟乙基淀粉和离心，以减少大量的红细胞和血浆，从而浓缩白细胞。根据存储系统的格式，该方法可以将同一冷冻空间中可存储的单元数量增加10倍。我们还研究了功能性干/祖细胞的比例，最初存在的实际上是提供给收件人时，解冻冷冻保存的PCB单位输注。当仍悬浮在高渗冷冻保存溶液中的解冻细胞与大量等渗溶液或血浆快速混合时，祖细胞活力可测量地降低。然而，由严重的溶质浓度梯度造成的渗透损伤可以通过解冻后简单的2倍稀释来避免，从而几乎完全恢复活的造血祖细胞。
Clinical evidence of hematopoietic restoration with placental/umbilical cord blood (PCB) grafts indicates that PCB can be a useful source of hematopoietic stem cells for routine bone marrow reconstitution. In the unrelated setting, human leukocyte antigen (HLA)-matched donors must be obtained for candidate patients and, hence, large panels of frozen HLA-typed PCB units must be established, The large volume of unprocessed units, consisting mostly of red blood cells, plasma, and cryopreservation medium, poses a serious difficulty in this effort because storage space in liquid nitrogen is limited and costly. We report here that almost all the hematopoietic colony-forming cells present in PCB units can be recovered in a uniform volume of 20 ml by using rouleaux formation induced by hydroxyethyl starch and centrifugation to reduce the bulk of erythrocytes and plasma and, thus, concentrate leukocytes. This method multiplies the number of units that can be stored in the same freezer space as much as 10-fold depending on the format of the storage system. We have also investigated the proportion of functional stem/progenitor cells initially present that are actually available to the recipient when thawed cryopreserved PCB units are infused. Progenitor cell viability is measurably decreased when thawed cells, still suspended in hypertonic cryopreservative solutions, are rapidly mixed with large volumes of isotonic Solutions or plasma. The osmotic damage inflicted by the severe solute concentration gradient, however, can be averted by a simple 2-fold dilution after thawing, providing almost total recovery of viable hematopoietic progenitor cells.