Unfixed CTC

ISET® enables lable-free enrichment of live CTC for further in vitro investigations and analyses.

At Rarecells, we understand that to study CTC, it is sometimes important to isolate them while still alive. This is possible with our own proprietary buffer and protocol. Fresh CTC can then be short-term cultured for analyses, including RNA analyses, or drug sensitivity tests. Please do not hesitate to contact us for more information. ISET® is so sensitive that it is capable of recovering one micropipetted cell in 10mL of blood.
Results submitted for publication, obtained by micropipetting in triplicate one fluorescent live A549 cell in 10mL of blood have shown the same unparalleled sensitivity obtained with fixed cells, i.e.  ISET® recovers the single micropipetted fresh cell in more than 90% of tests.
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Live CTC Workflow

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Live CTC Applications

After blood filtration, the live CTC are collected in suspension in 15 mL falcon tubes. Each cell resuspension tube can be analysed independently, therefore the Rarecells® System can allow multiplexing for live cells, as listed in the next paragraphs. As live CTC collection is a recently developed application of ISET®, some of the publications cited below do not report on ISET®.
In vitro CTC culture
Performances of the ISET® filtration process for live CTC collection were recently assessed thoroughly, demonstrating the unparalleled sensitivity of live cell collection (Laget et al. 2017 Plos One). Because ISET® allows enrichment of live cells with intact nucleus and cytoskeleton features, in vitro CTC culture is possible (Laget et al. 2017 Plos One).
CTC– derived Xenographt
Because ISET® allows enrichment of live cells with intact nucleus and cytoskeleton features, in vivo studies of CTC through generation of xenographts are possible (Laget et al. 2017 Plos One).CTC-derived xenographts have been proven to enable functional studies such as in vivo drug testing and to allow the generation of CTC-derived cell lines in vitro (Cayrefourcq et al. 2015 Cancer Res).
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Molecular analyses
Live cell collection by ISET® enables to uncover gene expression profiles of CTC and to study their transcriptomic heterogeneity by quantitative RT-PCR or by high throughput single-cell RNAseq .
Immunolabelling
Live CTC characterization by immunolabelling following ISET® filtration can guide the identification of specific CTC subsets since live cell collection by ISET® does not impact cell surface markers (Laget et al. 2017 Plos One).