Sato J, Kawano Y, Takaue Y, Hirao A, Makimoto A, Okamoto Y, Abe T, Shimokawa T, Iwai A, Kuroda Y
Department of Pediatrics, University Hospital of Tokushima, Japan.
Stem Cells. 1995 Sep;13(5):548-55. doi: 10.1002/stem.5530130513.
Some of the uncertainty regarding the use of cord blood (CB) in transplant settings includes the suspected relative rarity of hematopoietic stem cells (SC) in CB and the feasibility of incorporating a cell separation protocol to remove red blood cells, which may result in an unacceptable loss of SC. To address this uncertainty, we isolated a SC fraction by Percoll or Ficoll gradients from CB and peripheral blood (PB), which had been mobilized by chemotherapy. The frequencies of mononuclear cells (MNC), CD34+ cells, colony-forming units granulocyte-macrophage (CFU-GM), and the output of colony-forming cells (CFC) after five weeks in long-term culture (LTC) assay were then evaluated. The mean numbers of these cells per ml of CB sample before gradient separation were, respectively, 4.9 x 10(6), 13.8 x 10(4), 3.0 x 10(3) and 681 (n = 37). In the recovery phase of PB, these numbers were, respectively, 2.0 x 10(6), 14.9 x 10(4), 3.5 x 10(3) and 270 per ml of processed blood at apheresis (n = 35). After Percoll separation, the recovery rates of these cells were, respectively, 29%, 92%, 97% and 95% in CB, and 65%, 87%, 123% and 102% in PB. After Ficoll separation of CB, the rates were, respectively, 55%, 107%, 92% and 105%. These results suggest that CB contains an adequate number of more immature progenitors which can be retained after cell separation with Percoll or Ficoll, thereby making it feasible to incorporate a cell processing procedure into a CB transplant protocol. Percoll separation provided a greater enrichment of cells than Ficoll.
在移植环境中使用脐带血(CB)存在一些不确定性,包括怀疑CB中造血干细胞(SC)相对稀少,以及采用细胞分离方案去除红细胞的可行性,这可能导致SC不可接受的损失。为了解决这种不确定性,我们通过Percoll或Ficoll梯度从CB和经化疗动员的外周血(PB)中分离出SC组分。然后评估了长期培养(LTC)测定五周后单核细胞(MNC)、CD34+细胞、集落形成单位粒细胞-巨噬细胞(CFU-GM)的频率以及集落形成细胞(CFC)的产量。梯度分离前每毫升CB样品中这些细胞的平均数分别为4.9×10⁶、13.8×10⁴、3.0×10³和681(n = 37)。在PB的恢复期,每毫升单采处理血液中这些细胞的数量分别为2.0×10⁶、14.9×10⁴、3.5×10³和270(n = 35)。Percoll分离后,这些细胞在CB中的回收率分别为29%、92%、97%和95%,在PB中分别为65%、87%、123%和102%。CB经Ficoll分离后,回收率分别为55%、107%、92%和105%。这些结果表明,CB含有足够数量的更不成熟祖细胞,在用Percoll或Ficoll进行细胞分离后可以保留,从而使得将细胞处理程序纳入CB移植方案成为可能。Percoll分离比Ficoll能提供更大程度的细胞富集。