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Báo cáo sinh học: " Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành hóa học dành cho các bạn yêu hóa học tham khảo đề tài: Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients | Liu et al. Journal of Translational Medicine 2011 9 70 http www.translational-medicine.eom content 9 1 70 JOURNAL OF TRANSLATIONAL MEDICINE METHODOLOGY Open Access Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients 1 f 1 f 2 1 3 1 Zhian Liu Alberto Fusi Eva Klopocki Alexander Schmittel Ingeborg Tinhofer Anika Nonnenmacher and Ulrich Keilholz1 Abstract Background A limitation of positive selection strategies to enrich for circulating tumor cells CTCs is that there might be CTCs with insufficient expression of the surface target marker which may be missed by the procedure. We optimized a method for enrichment subsequent detection and characterization of CTCs based on depletion of the leukocyte fraction. Methods The 2-step protocol was developed for processing 20 mL blood and based on red blood cell lysis followed by leukocyte depletion. The remaining material was stained with the epithelial markers EpCAM and cytokeratin CK 7 8 or for the melanoma marker HMW-MAA MCSP. CTCs were detected by flow cytometry. CTCs enriched from blood of patients with carcinoma were defined as EpCAM CK CD45-. CTCs enriched from blood of patients with melanoma were defined as MCSP CD45-. One-hundred-sixteen consecutive blood samples from 70 patients with metastatic carcinomas n 48 or metastatic melanoma n 22 were analyzed. Results CTCs were detected in 47 of 84 blood samples 56 drawn from carcinoma patients and in 17 of 32 samples 53 from melanoma patients. CD45-EpCAM-CK was detected in pleural effusion specimens as well as in peripheral blood samples of patients with NSCLC. EpCAM-CK cells have been successfully cultured and passaged longer than six months suggesting their neoplastic origin. This was confirmed by CGH. By defining CTCs in carcinoma patients as CD45-CK and or EpCAM the detection rate increased to 73 61 84 . Conclusion Enriching CTCs using CD45 depletion allowed for detection of .