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通过免疫磁珠法和免疫吸附细胞分离技术纯化的CD34+骨髓细胞的比较。

Comparison of CD34+ bone marrow cells purified by immunomagnetic and immunoadsorption cell separation techniques.

作者信息

Firat H, Giarratana M C, Kobari L, Poloni A, Bouchet S, Labopin M, Gorin N C, Douay L

机构信息

Service d'Hématologie Biologique, Hôpital Armand Trousseau, Paris, France.

出版信息

Bone Marrow Transplant. 1998 May;21(9):933-8. doi: 10.1038/sj.bmt.1701200.

Abstract

We tested two positive selection techniques for separation of CD34+ cells from bone marrow and analyzed the yields of CD34+ cells, BFU-E, CFU-GM, CFU-MK and LTC-IC after selection and expansion. An immunoadsorption procedure (CellPro) and an immunomagnetic (Baxter) CD34+ cell separation method were employed to purify the same bone marrow samples from seven normal subjects. Mean yields of CFU-GM and CFU-MK and absolute numbers of LTC-ICs were not different in the two purified cell populations. In contrast, the mean recovery of BFU-E was significantly lower for the immunoadsorption (21 +/- 14%) than for the immunomagnetic technique (44 +/- 27%). After separation, CD34+ cells were evaluated in 10-day liquid cultures for their expansion capacity in terms of total cells and progenitors. The expansion capacity of progenitors such as CFU-GM, CFU-MK and especially BFU-E selected by immunoadsorption was higher than the capacity of progenitors obtained by immunomagnetism, although final total and progenitor cell numbers are similar. Our results suggest that the populations separated by the two techniques differ mainly in the expansion capacity of progenitors and in the recovery of BFU-E after the selection procedure. These differences between two methods, which already are widely employed in research and in clinical transplantation, should be taken into account when considering the aims of the experiments.

摘要

我们测试了两种从骨髓中分离CD34+细胞的阳性选择技术,并分析了选择和扩增后CD34+细胞、爆式红系集落形成单位(BFU-E)、粒系巨噬系集落形成单位(CFU-GM)、巨核系集落形成单位(CFU-MK)和长期培养起始细胞(LTC-IC)的产量。采用免疫吸附程序(CellPro)和免疫磁珠法(百特公司)从7名正常受试者的相同骨髓样本中纯化细胞。两种纯化细胞群体中CFU-GM和CFU-MK的平均产量以及LTC-IC的绝对数量没有差异。相比之下,免疫吸附法的BFU-E平均回收率(21±14%)显著低于免疫磁珠技术(44±27%)。分离后,对CD34+细胞进行为期10天的液体培养,评估其在总细胞和祖细胞方面的扩增能力。尽管最终的总细胞数和祖细胞数相似,但免疫吸附法选择的祖细胞如CFU-GM、CFU-MK尤其是BFU-E的扩增能力高于免疫磁珠法获得的祖细胞。我们的结果表明,两种技术分离的细胞群体主要在祖细胞的扩增能力和选择程序后BFU-E的回收率方面存在差异。在考虑实验目的时,应考虑这两种已广泛应用于研究和临床移植的方法之间的这些差异。

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