Van Epps D E, Bender J, Lee W, Schilling M, Smith A, Smith S, Unverzagt K, Law P, Burgess J
Immunotherapy Division, Baxter Healthcare Corporation, Round Lake, Illinois 60073, USA.
Blood Cells. 1994;20(2-3):411-23.
Stem and progenitor cells from a variety of sources including bone marrow, cord blood, and peripheral blood have been used for transplantation. This study compares CD34 cells from all three sources. Flow cytometry analysis of CD34 cells in multiple samples of normal peripheral blood and patient peripheral blood mobilized with chemotherapy (cyclophosphamide/VP16), chemotherapy plus granulocyte colony stimulating factor (G-CSF), and G-CSF alone were compared to bone marrow and cord blood. Although the relative distribution of CD34 percentages in each preparation of cells varied widely, on average the percentage of CD34 cells in these different preparations was 0.15%, 0.6%, 2%, 0.45%, 1.68%, and 0.83% respectively. CD34 subset analysis was performed on these cell preparations using multicolor flow cytometry and antibodies to CD33, CD13, CD45RA, CD19, CD71, and CD38. The major differences observed were that bone marrow CD34 cells contain high percentages of CD19+ cells not found in significant quantity in the other cell preparations and cord blood CD34 cells contained a higher percentage of CD38-cells than the other cell preparations. A magnetic bead system was used with anti-CD34 antibody to purify CD34 cells from mobilized peripheral blood apheresis products, cord blood, and bone marrow. Efficient selection with high purities of CD34 cells was achieved with each of the cell preparations. Comparison of colony-forming activity of each of the cell preparations showed cord blood and mobilized peripheral blood to have slightly higher cloning efficiencies than bone marrow with higher numbers of erythroid blast-forming units (BFU-E) also observed in cord blood CD34 cells. Culture of isolated CD34 cells in liquid culture with interleukin-3, stem cell factor, G-CSF, and granulocyte-macrophage GM-CSF showed over a 100-fold expansion in cell numbers after 25 days, with the peak expansion of colony-forming cells occurring between days 11 and 16. Analysis of day-10 cells from these cultures showed them to be predominantly promyelocytes, myelocytes, and metamyelocytes, with cord blood CD34 cultures showing more promyelocytes than peripheral blood or bone marrow and bone marrow showing more metamyelocytes. Comparison of the proliferation of CD34 cells from these different cell preparations showed that cord blood CD34 cells cultured for 10 days averaged an 85-fold increase in cell numbers followed by mobilized peripheral blood CD34 cells, with an average 56-fold increase, and bone marrow CD34 cells, with an average 49-fold increase.
包括骨髓、脐带血和外周血在内的多种来源的干细胞和祖细胞已被用于移植。本研究比较了来自所有这三种来源的CD34细胞。对正常外周血和经化疗（环磷酰胺/VP16）、化疗加粒细胞集落刺激因子（G-CSF）以及单独使用G-CSF动员的患者外周血的多个样本中的CD34细胞进行流式细胞术分析，并与骨髓和脐带血进行比较。尽管每种细胞制剂中CD34百分比的相对分布差异很大，但这些不同制剂中CD34细胞的百分比平均分别为0.15%、0.6%、2%、0.45%、1.68%和0.83%。使用多色流式细胞术和针对CD33、CD13、CD45RA、CD19、CD71和CD38的抗体对这些细胞制剂进行CD34亚群分析。观察到的主要差异是骨髓CD34细胞中含有高百分比的CD19+细胞，而在其他细胞制剂中未大量发现，并且脐带血CD34细胞中CD38-细胞的百分比高于其他细胞制剂。使用抗CD34抗体的磁珠系统从动员的外周血单采产品、脐带血和骨髓中纯化CD34细胞。每种细胞制剂都实现了对CD34细胞的高效选择且纯度高。对每种细胞制剂的集落形成活性进行比较，结果显示脐带血和动员的外周血的克隆效率略高于骨髓，在脐带血CD34细胞中还观察到更高数量的红系爆式集落形成单位（BFU-E）。将分离的CD34细胞在含有白细胞介素-3、干细胞因子、G-CSF和粒细胞-巨噬细胞集落刺激因子（GM-CSF）的液体培养基中培养，25天后细胞数量扩增超过100倍，集落形成细胞的峰值扩增发生在第11天至16天之间。对这些培养物第10天的细胞进行分析，结果显示它们主要是早幼粒细胞、中幼粒细胞和晚幼粒细胞，脐带血CD34培养物中的早幼粒细胞比外周血或骨髓中的更多，而骨髓中的晚幼粒细胞更多。对来自这些不同细胞制剂的CD34细胞的增殖进行比较，结果显示培养10天的脐带血CD34细胞数量平均增加85倍，其次是动员的外周血CD34细胞，平均增加56倍，骨髓CD34细胞平均增加49倍。
