Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cells

作者：Liying Yang, James C. Lang, Priya Balasubramanian, Kris R. Jatana, David Schuller, Amit Agrawal, Maciej Zborowski, Jeffrey J. Chalmers · 发表于：Biotechnology and Bioengineering · 年份：2008 · DOI：10.1002/bit.22066 · 被引用次数：200 · 研究领域：Cancer Cells and Metastasis、Mesenchymal stem cell research、Cancer Genomics and Diagnostics

The optimization of a purely negative depletion, enrichment process for circulating tumor cells (CTCs) 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 x 10(9) to 8.04 x 10(3) 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. Since the initial number of CTC in a patient's blood is unknown, and most probably varies from patient to patient, the recovery of the CTC is unknown. However, spiking studies of a cancer cell line into normal blood, and subsequent enrichment using the optimized protocol indicated an average recovery of approximately 83%. Unlike a majority of other publishe...