Fast isolation and ex vivo culture of circulating tumor cells from the peripheral blood of lung cancer patients.
Yi Chuan
; 39(1): 66-74, 2017 01 20.
Article
em En
| MEDLINE
| ID: mdl-28115307
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 EGFRãKRASãPIK3CAãTP53 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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Técnicas de Cultura de Células
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Citometria de Fluxo
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Neoplasias Pulmonares
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Células Neoplásicas Circulantes
Limite:
Humans
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article