Confocal Images of Circulating Tumor Cells Obtained Using a Methodology and Technology That Removes Normal Cells
Confocal Images of Circulating Tumor Cells Obtained Using a Methodology and Technology That Removes Normal Cells
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DOI：
10.1021/mp9000519
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发表时间：
2009-09-01
影响因子：
4.9
通讯作者：
Chalmers, Jeffrey J.
中科院分区：
文献类型：
--
作者：
Balasubramanian, Priya;Yang, Liying;Chalmers, Jeffrey J.
采用全负富集技术检测头颈癌患者外周血循环肿瘤细胞ctc。在32份血液样本中，63%含有CTCs，每毫升血液中鉴定的CTCs数量从0到214不等。最终纯度从9个细胞中的1个CTC到2万个细胞中的1个CTC不等，最终纯度是CTC数量和特异性富集性能的函数。与先前的报道一致，如果：(1)基于DAPI染色的CTC含有细胞核，(2)细胞角蛋白染色阳性，(3)核与细胞质比高，则CTC被阳性鉴定。此外，对于被认为ctc阳性的血液样本，富集后的样本必须通过RT-PCR检测上皮生长因子受体（EGFR）呈阳性。虽然大多数血液样本是在手术前和在手术前采集的，但在所有术前和术后配对样本中，检测到大量的ctc。除了传统的宽视场荧光显微镜下的显微观察外，还在共聚焦显微镜下观察了这些富集样品。正如预期的那样，FITC染色的细胞角蛋白出现在细胞质中，这些阳性染色的细胞在细胞自旋上的平均大小在8-12 μ m之间。未来的研究将涉及调查这些CTC上是否存在癌症干细胞和间质标志物，以及患者预后与存在的CTC数量和类型的相关性。
A completely negative enrichment technology was used to detect circulating tumor cells, CTCs, in the peripheral blood of head and neck cancer patients. Of 32 blood samples, 63% contained CTCs and the number of CTCs identified per mL of blood collected ranged from 0 to 214. The final purity ranged from 1 CTC in 9 total cells to 1 CTC in 20,000 total cells, the final purity being both a function of the number of CTCs and the performance of the specific enrichment. Consistent with previous reports, CTC were positively identified if: (1) they contained a nucleus based on DAPI stain, (2) stained positive for cytokeratins, and (3) have a high nuclei to cytoplasmic ratio. In addition, for a blood sample to be considered positive for CTCs, the enriched sample must be positive for epithelial growth factor receptor, EGFR, as measured by RT-PCR. While most of the blood samples were obtained during surgery, a number were taken prior to and during surgery, In all of the pre- and postsurgery paired samples, significant numbers of CTCs were detected. A number of these enriched samples were observed under confocal microscope in addition to the microscopic observations under traditional wide-field fluorescent microscope. As expected, the FITC stained cytokeratins appeared in the cytoplasm and the average size of these positively stained cells, on the cytospin, was in the range of 8-12 mu m. Future studies will involve the investigation if cancer stem cell and mesenchymal markers are present on these CTCs and correlations of patient outcome to the number and type of CTC present.