Silvestri F, Banavali S, Yin M, Gopal V, Baccarani M, Preisler H D
University of Cincinnati Medical Center, OH.
Leuk Lymphoma. 1992 Nov;8(4-5):389-96. doi: 10.3109/10428199209051019.
The aim of the present study was to compare two different methods for obtaining CD34+ cells from the peripheral blood or the bone marrow of patients with acute non-lymphocytic leukemia (ANLL). Twenty-two samples, obtained from 19 patients, were density cut (Ficoll-Hypaque 1.077) and, after incubation with My10 antibody, separated by panning or by immunomagnetic beads. Immunomagnetic beads provided a significantly better separation than panning, either in terms of concentration of CD34+ cells (85.5 +/- 11.6% vs. 55.7 +/- 25.0%, p = 0.003) or in terms of depletion of CD34+ cells (3.9 +/- 8.0% vs. 30.9 +/- 26.3%, p = 0.008). This was consistent with the virtually complete depletion of colony forming cells (CFC) in the CD34 negative fraction and the recovery of virtually all the CFC in the positive fraction in the samples separated by immunomagnetic beads. In conclusion, separation by immunomagnetic beads can allow collection of nearly pure CD34+ and CD34- cell populations from patients with ANLL, thereby facilitating the study of the biological characteristics of these cell populations. Moreover the method is less time consuming than panning and is not toxic to the CFC.
本研究的目的是比较两种从急性非淋巴细胞白血病(ANLL)患者外周血或骨髓中获取CD34+细胞的不同方法。从19例患者中获取的22个样本进行密度梯度离心(Ficoll-Hypaque 1.077),与My10抗体孵育后,通过淘选法或免疫磁珠法进行分离。无论是从CD34+细胞浓度(85.5±11.6%对55.7±25.0%,p = 0.003)还是从CD34+细胞的去除率(3.9±8.0%对30.9±26.3%,p = 0.008)来看,免疫磁珠法的分离效果都明显优于淘选法。这与免疫磁珠法分离的样本中CD34阴性部分的集落形成细胞(CFC)几乎完全去除以及阳性部分几乎所有CFC的回收情况一致。总之,免疫磁珠法分离能够从ANLL患者中收集到几乎纯的CD34+和CD34-细胞群体,从而便于对这些细胞群体的生物学特性进行研究。此外,该方法比淘选法耗时少,且对CFC无毒害作用。