Isolation and detection of circulating tumour cells from metastatic melanoma patients using a slanted spiral microfluidic device.
Isolation and detection of circulating tumour cells from metastatic melanoma patients using a slanted spiral microfluidic device.
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DOI：
10.18632/oncotarget.18641
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发表时间：
2017-09-15
期刊：
影响因子：
--
通讯作者：
Ziman M
中科院分区：
文献类型：
--
作者：
Aya-Bonilla CA;Marsavela G;Freeman JB;Lomma C;Frank MH;Khattak MA;Meniawy TM;Millward M;Warkiani ME;Gray ES;Ziman M
循环肿瘤细胞（CTC）是有前途的癌症生物标志物。已经开发了几种方法来从血液样品中分离CTC。然而，由于黑色素瘤CTC的特殊异质性，其分离非常具有挑战性，这阻碍了其生物学和临床研究。因此，在黑素瘤中非常需要基于CTC的物理性质而不是表面标志物表达来分离CTC的方法，例如微流体装置。在这里，我们评估了倾斜螺旋微流体装置通过无标记富集分离黑素瘤CTC的能力。我们证明，该装置在一轮或两轮富集后，分别产生超过80%和55%的加标黑色素瘤细胞的回收率。同时，分别用一轮或两轮富集实现了白色血细胞的2至3个对数减少。我们使用多标记流式细胞术、免疫细胞化学和基因表达来表征分离的CTC。结果表明，来自转移性黑色素瘤患者的CTC是高度异质性的，并且通常表达干细胞样标志物，如PAX3和ABCB 5。用于黑色素瘤CTC分离的倾斜微流体装置的实施使得能够进一步理解用于生物标志物开发的黑色素瘤转移的生物学，并为未来的治疗方法提供信息。
Circulating Tumour Cells (CTCs) are promising cancer biomarkers. Several methods have been developed to isolate CTCs from blood samples. However, the isolation of melanoma CTCs is very challenging as a result of their extraordinary heterogeneity, which has hindered their biological and clinical study. Thus, methods that isolate CTCs based on their physical properties, rather than surface marker expression, such as microfluidic devices, are greatly needed in melanoma. Here, we assessed the ability of the slanted spiral microfluidic device to isolate melanoma CTCs via label-free enrichment. We demonstrated that this device yields recovery rates of spiked melanoma cells of over 80% and 55%, after one or two rounds of enrichment, respectively. Concurrently, a two to three log reduction of white blood cells was achieved with one or two rounds of enrichment, respectively. We characterised the isolated CTCs using multimarker flow cytometry, immunocytochemistry and gene expression. The results demonstrated that CTCs from metastatic melanoma patients were highly heterogeneous and commonly expressed stem-like markers such as PAX3 and ABCB5. The implementation of the slanted microfluidic device for melanoma CTC isolation enables further understanding of the biology of melanoma metastasis for biomarker development and to inform future treatment approaches.
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影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
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影响因子：
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作者：
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通讯作者：
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