A new automated cell washer device for thawed cord blood units
A new automated cell washer device for thawed cord blood units
复制标题
DOI：
10.1111/j.1537-2995.2004.03389.x
复制
发表时间：
2004-06-01
期刊：
影响因子：
2.9
通讯作者：
Rocchi, L
中科院分区：
文献类型：
--
作者：
Perotti, CG;Del Fante, C;Rocchi, L
背景：现有的从解冻的脐带血(UCB)中清洗DMSO的技术基本上是基于开放系统中的标准离心法，其中存在不同程度的细胞损失。研究设计和方法：我们评估了一种新的自动封闭式设备(Cytomate，Baxter，IL)从解冻的脐带血中清洗DMSO的能力，同时节省了CD34+细胞和MNC的祖细胞和附属细胞。我们修改了该装置的标准软件，并计算了25个脐带血单位的细胞回收率。结果：为了评价Cytomate装置的效率，我们考虑了解冻(洗涤前)和洗涤后细胞恢复的情况。有核细胞总数的平均回收率(%)为63.30(40.12-89.00)，CD34+细胞为70.20(11.51-89.01)，CD3+细胞为61.01(28.80-87.08)，CD4+细胞为62.53(30.62-96.73)，CD8+细胞为57.4(26.87-94.72)，CD19+细胞为63.33(39.10-90.33)，CD16+/56+细胞为70.67(范围，CFU-GM为74.33(20.23-98.60)，CFU-GM为82.34(14.83-247.12)，存活率为89.67(70.74-98.30)。总工作时间7~20分钟，平均15分钟。结论：Cytomate装置在细胞回收和维持脐血移植物的克隆形成能力方面表现出令人满意的效率。DMSO的去除几乎是完全的，对接受者来说具有明显的优势。最后，在封闭系统中进行的整个操作被证明是安全的，保持了移植物的无菌。
BACKGROUND: The current available techniques to wash out DMSO from thawed umbilical cord blood (UCB) units are based essentially on standard centrifugation in an open system with various degrees of cell loss.STUDY DESIGN AND METHODS: We evaluated the capacity of a new automated closed device (Cytomate, Baxter, IL) to wash out the DMSO from thawed UCB units, saving at the same time the progenitor and accessory cells in terms of CD34+ cells and MNCs. We modified the standard software of the device and calculated the cell recovery on 25 UCB units. Moreover, we set up a new gas chromatographic method to exactly detect the DMSO removal rate.RESULTS: To evaluate the efficiency of the Cytomate device, we considered the postthawing (prewashing) versus postwashing cell recovery. The average recovery (%) in terms of total nucleated cells was 63.30 (range, 40.12-89.00), CD34+ cells was 70.20 (range, 11.51-89.01), CD3+ cells was 61.01 (range, 28.80-87.08), CD4+ cells was 62.53 (range, 30.62-96.73), CD8+ cells was 57.4 (range, 26.87-94.72), CD19+ cells was 63.33 (range, 39.10-90.33), CD16+/56+ cells was 70.67 (range, 8.91-98.40), CFU-GM was 74.33 (range, 20.23-98.60), total CFUs was 82.34 (range, 14.83-247.12), and viability was 89.67(range, 70.74-98.30). The total working time required was, on average, 15 minutes (range, 7-20).CONCLUSIONS: The Cytomate device demonstrated a satisfying efficiency in cell recovery and in maintaining the clonogenic power of the UCB graft. The removal rate of DMSO was practically complete with evident advantages for the recipient. Finally, the entire manipulation performed in a closed system revealed to be safe, maintaining the sterility of the graft.