Han Xiaohong, Shi Yuankai, Zhang Wei
Department of Medical Oncology, Cancer Institute (Hospital), Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.
Zhonghua Yi Xue Za Zhi. 2002 Feb 10;82(3):203-6.
To evaluate the effect of immunomagnetic technique in purging of peripheral blood progenitor cell (PBPC) of breast cancer patients.
A sample of APBSC from non-breast cancer patients mixed with 1% MCF-7 cell lines and 10 samples of APBSC from breast cancer patients were subjected to positive purging, negative purging, and positive/positive purging. The CD34 + cell purity, CD34 + cell recovery rate and the enrichment of colony form units were compared. The number of tumor cells before and after immunochemistry (ICC) and flow detected purging procedure cytometry (FCM). The influence of refrigeration on the effect of purging was analyzed.
The TC depletion in 10 samples of PBPC of breast cancer patients after positive purging was 2.1 (1.1 approximately 2.7) log steps, the CD34 + cell purity was 55.1 (25.9 approximately 99.5)%, and the CD34 + cell recovery was 39.4 (34.0 approximately 80.0)%. After the 6 samples of ICC positive PBPC from breast cancer patients were subjected to negative selection and subsequent positive purging, the mean CD34 cell purity was 64.3 (34.0 approximately 86.4)%, the mean CD34 + cell recovery was 35.0 (24.0 approximately 52.5)%, the purging efficiency was 3.0 (1.8 approximately 3.9) log, and the enrichment of colony-forming units-granulocyte macrophage (CFU-GM) and burst-forming unit-erythrocyte (BFU-E) were 2.2 and 3.1 respectively. TCs were detectable in all 6 cases prior to the purging procedure by ICC and FCM. After the -/+ purging procedure, only one case was tumor cell positive by ICC and residual tumor cells could be detected by FCM, however the number of TCs was evidently decreased. Along with the extension of refrigeration time, the CD34 cell purity, CD34 cell recovery, and tumor cell clearance rate decreased.
Immunomagnetic double negative/positive selections an effective, rapid and simple purging method.
评估免疫磁珠技术对乳腺癌患者外周血祖细胞(PBPC)的净化效果。
将非乳腺癌患者的一份采集的外周血干细胞(APBSC)样本与1%的MCF - 7细胞系混合,以及10份乳腺癌患者的APBSC样本分别进行阳性净化、阴性净化和阳性/阳性净化。比较CD34⁺细胞纯度、CD34⁺细胞回收率以及集落形成单位的富集情况。通过免疫化学(ICC)和流式细胞术(FCM)检测净化前后肿瘤细胞的数量。分析冷藏对净化效果的影响。
10份乳腺癌患者PBPC样本经阳性净化后,肿瘤细胞清除率为2.1(1.1至2.7)log级,CD34⁺细胞纯度为55.1(25.9至99.5)%,CD34⁺细胞回收率为39.4(34.0至80.0)%。6份ICC阳性的乳腺癌患者PBPC样本经阴性选择及随后的阳性净化后,平均CD34细胞纯度为64.3(34.0至86.4)%,平均CD34⁺细胞回收率为35.0(24.0至52.5)%,净化效率为3.0(1.8至3.9)log,粒细胞巨噬细胞集落形成单位(CFU - GM)和红细胞爆式集落形成单位(BFU - E)的富集倍数分别为2.2和3.1。在净化前,所有6例样本通过ICC和FCM均可检测到肿瘤细胞。经过阴性/阳性净化后,仅1例通过ICC检测为肿瘤细胞阳性,通过FCM可检测到残留肿瘤细胞,但肿瘤细胞数量明显减少。随着冷藏时间延长,CD34细胞纯度、CD34细胞回收率及肿瘤细胞清除率均下降。
免疫磁珠双阴性/阳性选择是一种有效、快速且简便的净化方法。