Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cells.
Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cells.
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DOI：
10.1002/bit.22066
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发表时间：
2009-02-01
影响因子：
3.8
通讯作者：
Chalmers, Jeffrey J.
中科院分区：
文献类型：
--
作者：
Yang, Liying;Lang, James C.;Balasubramanian, Priya;Jatana, Kris R.;Schuller, David;Agrawal, Amit;Zborowski, Maciej;Chalmers, Jeffrey J.
关键词：
提出了头颈癌患者外周血中循环肿瘤细胞CTC的纯阴性耗竭、富集过程的优化。富集过程使用红细胞裂解步骤，然后进行免疫磁性标记，随后去除CD 45阳性细胞。许多相关变量被量化或试图量化，这些变量控制着富集过程的性能。评估了六种不同的免疫磁性标记组合以及相对于血液来源的性能的显著差异：从红十字会购买的血沉棕黄层，来自正常供体的新鲜外周血和来自人类癌症患者的新鲜外周血。优化后，该过程能够将癌症患者血液中的正常血细胞数量从4.05 × 109减少到8.04 × 103个细胞/ml，并且平均每ml血液仍能回收2.32 CTC。对于检测到CTC的所有测试的癌症患者血液样品（26名患者中的20名），CTC的平均回收率为21.7/ml血液，范围为282至0.53 CTC/ml血液。与大多数其他已发表的研究不同，本研究的重点是量化尽可能多的因素，以促进过程的优化，并为未来的性能比较提供信息。作者不知道有任何其他报告的研究在纯负富集操作模式下实现了此处报告的性能（5.76 log 10）。富集过程的这种操作模式提供了显著的灵活性，因为它对于定义CTC的属性没有偏见;从而允许研究人员或临床医生使用他们选择的任何标记物来定义最终的富集产物是否含有CTC或与特定问题相关的其他细胞类型（即CTC主要具有上皮细胞或间充质细胞特征？）。
The optimization of a purely negative depletion, enrichment process for circulating tumor cells, CTC's, in the peripheral blood of Head and Neck cancer patients is presented. The enrichment process uses a red cell lysis step followed by immunomagnetic labeling, and subsequent depletion, of CD45 positive cells. A number of relevant variables are quantified, or attempted to be quantified, which control the performance of the enrichment process. Six different immunomagnetic labeling combinations were evaluated as well as the significant difference in performance with respect to the blood source: buffy coats purchased from the Red Cross, fresh, peripheral blood from normal donors, and fresh peripheral blood from human cancer patients. After optimization, the process is able to reduce the number of normal blood cells in a cancer patient's blood from 4.05 × 109 to 8.04 × 103 cells/ml and still recover, on average, 2.32 CTC per ml of blood. For all of the cancer patient blood samples tested in which CTC were detected (20 out of 26 patients) the average recovery of CTCs was 21.7 per ml of blood, with a range of 282 to 0.53 CTC per ml of blood. Unlike a majority of other published studies, this study focused on quantifying as many factors as possible to facilitate both the optimization of the process as well as provide information for future performance comparisons. The authors are not aware any other reported study which has achieved the performance reported here (a 5.76 log10) in a purely negative enrichment mode of operation. Such a mode of operation of an enrichment process provides significant flexibility in that it has no bias with respect to what attributes define a CTC; thereby allowing the researcher or clinician to use any maker they choose to define whether the final, enrich product contains CTC's or other cell type relevant to the specific question (i.e. does the CTC have predominately epithelia or mesenchymal characteristics?).
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影响因子：
11.4
作者：
RALPH, SJ;THOMAS, ML;TROWBRIDGE, IS
通讯作者：
TROWBRIDGE, IS
影响因子：
2.7
作者：
Kokkinos, Maria I.;Wafai, Razan;Waltham, Mark
通讯作者：
Waltham, Mark
影响因子：
2.7
作者：
Chaffer, Christine L.;Thompson, Erik W.;Williams, Elizabeth D.
通讯作者：
Williams, Elizabeth D.
影响因子：
11.5
作者：
Riethdorf, Sabine;Fritsche, Herbert;Pantel, Klaus
通讯作者：
Pantel, Klaus
DOI：
10.1073/pnas.95.8.4589
发表时间：
1998-04-14
影响因子：
11.1
作者：
Racila, E;Euhus, D;Uhr, JW
通讯作者：
Uhr, JW