Fast isolation and<i> ex vivo </i>culture of circulating tumor cells from the peripheral blood of lung cancer patients

Wenjun Wu, Zhihua Wang, Zhuo Wang, Yuliang Deng, Qihui Shi

doi:10.16288/j.yczz.16-217

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Hereditas(Beijing) >

2017 , Vol. 39 >Issue 1: 66 - 74

DOI: https://doi.org/10.16288/j.yczz.16-217

Fast isolation and ex vivo culture of circulating tumor cells from the peripheral blood of lung cancer patients

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Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University, Shanghai 200240, China

Received date: 2016-06-17

Revised date: 2016-10-19

Online published: 2017-12-24

Supported by

the National Natural Science Foundation of China(81371712,81501613);the Natural Science Foundation of Shanghai(13ZR1456100);Research Fund for the Docoral Program of High Education of China(20130073120106)

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Abstract

Circulating tumor cells (CTCs) are free tumor cells shed from tumor site and enter into blood circulation. CTCs represent a reliable source of tumor cells for the molecular characteristics of the original tumor. However, the extraordinary rarity of CTCs makes the subsequent molecular and functional analysis technically challenging. Here, we describe a one-step microfludics-based immunomagnetic isolation method to isolate CTCs directly from the whole blood of lung adenocarcinoma patients. This method avoids harsh sample preparation and enrichment steps, and therefore preserves the viability (>90%) of CTCs during the in vitro isolation. The isolated CTCs are enriched in small volume (80 μL) and cultured ex vivo that leads to successful ex vivo expansion. The expanded CTCs can be frozen and thawed, which shows cell line property. Genetic sequencing on EGFR、KRAS、PIK3CA、TP53 and BRAF and metabolic assay (2-NBDG) are utilized to characterize the expanded CTCs. Our results demostrated that this method is suitable for ex vivo expansion of CTCs facilitates. The genomic, proteomic and metabolic analyses of CTCs have guiding significance in tumor precise treatment.

Key words： circulating tumor cells; microfluidics; ex vivo culture; liquid biospy

Cite this article

Wenjun Wu, Zhihua Wang, Zhuo Wang, Yuliang Deng, Qihui Shi . Fast isolation and ex vivo culture of circulating tumor cells from the peripheral blood of lung cancer patients[J]. Hereditas(Beijing), 2017 , 39(1) : 66 -74 . DOI: 10.16288/j.yczz.16-217

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