Panterne B, Richard M-J, Sabatini C, Pouthier F, Mouillot L, Bardey D, Boulanger F, Créa S, Dal Cortivo L, Decot V, Fleury-Cappellesso S, Giraud C, Lapierre V, Léauté A-G, Le Berre C, Lemarié C, Piard N, Rapatel C, Rosenzwajg M
Afssaps, DLC, unité produits sanguins et thérapie cellulaire, 143/147, boulevard Anatole-France, 93285 St-Denis cedex, France.
Transfus Clin Biol. 2010 Apr;17(2):41-6. doi: 10.1016/j.tracli.2010.06.006. Epub 2010 Jul 31.
Today, haematopoietic stem cell graft from placental blood concerns more than 15 % of allogeneic grafts. An inter-laboratory study of the quality control of defrosted cord blood units has been coordinated by the French society for cell and tissue bioengineering (SFBCT), with the cord blood bank of Bourgogne Franche-Comté and controlled by the French health products safety agency (Afssaps). The aim of this study is to ensure the inter-laboratory reproducibility of the quality controls practised by the banks during defrosting. The cellular outputs were analyzed according to the defrosting techniques, according to the method used in flow cytometry: single-platform (SP) versus double-platform (DP), or the product nature, i.e. in total blood or miniaturized.
Forty-two units of placental blood (USP), which were out of range were provided for defrosting to 14 participating sites. USP were defrosted and controlled according to the procedures of each bank. Once the USP is defrosted, a part of the product was controlled by the site and the other part by Afssaps. Following controls were carried out: numeration of the total nucleated cells (TNC) and of CD34+ cells (made by a SP method in Afssaps) and functional assay.
Concerning TNC, the defrosting sites obtained a cellular output of 94 %+/-28 in day 0 compared with an output of 72 %+/-24 in Afssaps showing a rather good stability of the USP transmitted with an average deviation of 23 %+/-22. The freezing process with or without reduction of volume does not affect this variation. Concerning the numeration of CD34+ cells, the average deviation between the participating sites and Afssaps was 29 %+/-23 compared with 21 %+/-16 for the sites using a SP method against 47 %+/-25 for those using a DP method. The CD34+ outputs are equal to 82 % +/- 60 in day 0 for the participating sites against 52 %+/-20 for Afssaps. For the sites using a DP method, it is stressed that this output is particularly high with a rate of 126 %+/-90 (n=15) whereas it is 62 %+/-20 (n=32) for the sites using a SP method.
These results underline a good stability of viable CD34+ cells and a greater reliability of the SP methods for the CD34+ cell numeration for these defrosted USP. Lastly, the results of the functional assay regarding the average clonogenicities (equal to 15 %) reinforce the conclusions on the quality of the defrosted products.
如今，胎盘血造血干细胞移植占所有异体移植的比例超过15%。法国细胞与组织生物工程学会（SFBCT）协调开展了一项关于解冻脐带血单位质量控制的实验室间研究，该研究由勃艮第弗朗什 - 孔泰脐带血库进行，并由法国健康产品安全局（Afssaps）监管。本研究的目的是确保各血库在解冻过程中所进行的质量控制在实验室间具有可重复性。根据解冻技术、流式细胞术所采用的方法（单平台[SP]与双平台[DP]）或产品性质（即全血或小型化产品）对细胞产出进行分析。
向14个参与研究的机构提供了42个超出范围的胎盘血单位（USP）用于解冻。USP按照各血库的程序进行解冻和检测。USP解冻后，一部分产品由各机构检测，另一部分由Afssaps检测。进行了以下检测：总核细胞（TNC）计数、CD34 +细胞计数（Afssaps采用SP方法）以及功能检测。
关于TNC，解冻当天各解冻机构的细胞产出为94%±28，而Afssaps的产出为72%±24，表明所传递的USP具有相当好的稳定性，平均偏差为23%±22。有无体积缩减的冷冻过程均不影响这种变化。关于CD34 +细胞计数，参与研究的机构与Afssaps之间的平均偏差为29%±23，而采用SP方法的机构之间的平均偏差为21%±16，采用DP方法的机构之间的平均偏差为47%±25。参与研究的机构在解冻当天CD34 +细胞产出为82%±60，而Afssaps为52%±20。对于采用DP方法的机构，需强调该产出特别高，比率为126%±90（n = 15），而采用SP方法的机构该比率为62%±20（n = 32）。
这些结果突显了解冻后USP中存活CD34 +细胞具有良好的稳定性，以及SP方法在这些解冻USP的CD34 +细胞计数方面具有更高的可靠性。最后，关于平均克隆形成能力（等于15%）的功能检测结果强化了关于解冻产品质量的结论。
