Scanning Electron Microscopy of Circulating Tumor Cells and Tumor-Derived Extracellular Vesicles.
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Embargo End Date
ICR Authors
Authors
Nanou, A
Crespo, M
Flohr, P
De Bono, JS
Terstappen, LWMM
Crespo, M
Flohr, P
De Bono, JS
Terstappen, LWMM
Document Type
Journal Article
Date
2018-10-31
Date Accepted
2018-10-30
Abstract
To explore morphological features of circulating tumor cells (CTCs) and tumor-derived extracellular vesicles (tdEVs), we developed a protocol for scanning electron microscopy (SEM) of CTCs and tdEVs. CTCs and tdEVs were isolated by immunomagnetic enrichment based on their Epithelial Cell Adhesion Molecule (EpCAM) expression or by physical separation through 5 μm microsieves from 7.5 mL of blood from Castration-Resistant Prostate Cancer (CRPC) patients. Protocols were optimized using blood samples of healthy donors spiked with PC3 and LNCaP cell lines. CTCs and tdEVs were identified among the enriched cells by fluorescence microscopy. The positions of DNA+, CK+, CD45- CTCs and DNA-, CK+, CD45- tdEVs on the CellSearch cartridges and microsieves were recorded. After gradual dehydration and chemical drying, the regions of interest were imaged by SEM. CellSearch CTCs retained their morphology revealing various shapes, some of which were clearly associated with CTCs undergoing apoptosis. The ferrofluid was clearly distinguishable, shielding major portions of all isolated objects. CTCs and leukocytes on microsieves were clearly visible, but revealed physical damage attributed to the physical forces that cells exhibit while entering one or multiple pores. tdEVs could not be identified on the microsieves as they passed through the pores. Insights on the underlying mechanism of each isolation technique could be obtained. Complete detailed morphological characteristics of CTCs are, however, masked by both techniques.
Citation
Cancers, 2018, 10 (11)
Source Title
Publisher
MDPI
ISSN
2072-6694
eISSN
2072-6694
Collections
Research Team
Cancer Biomarkers
Prostate Cancer Targeted Therapy Group
Prostate Cancer Targeted Therapy Group