Sensitive and Specific Biomimetic Lipid Coated Microfluidics to Isolate Viable Circulating Tumor Cells and Microemboli for Cancer Detection

Abstract

Here we presented a simple and effective membrane mimetic microfluidic device with antibody conjugated supported lipid bilayer (SLB) “smart coating” to capture viable circulating tumor cells (CTCs) and circulating tumor microemboli (CTM) directly from whole blood of all stage clinical cancer patients. The non-covalently bound SLB was able to promote dynamic clustering of lipid-tethered antibodies to CTC antigens and minimized non-specific blood cells retention through its non-fouling nature. A gentle flow further flushed away loosely-bound blood cells to achieve high purity of CTCs, and a stream of air foam injected disintegrate the SLB assemblies to release intact and viable CTCs from the chip. Human blood spiked cancer cell line test showed the ~95% overall efficiency to recover both CTCs and CTMs. Live/dead assay showed that at least 86% of recovered cells maintain viability. By using 2 mL of peripheral blood, the CTCs and CTMs counts of 63 healthy and colorectal cancer donors were positively correlated with the cancer progression. In summary, a simple and effective strategy utilizing biomimetic principle was developed to retrieve viable CTCs for enumeration, molecular analysis, as well as ex vivo culture over weeks. Due to the high sensitivity and specificity, it is the first time to show the high detection rates and quantity of CTCs in non-metastatic cancer patients. This work offers the values in both early cancer detection and prognosis of CTC and provides an accurate non-invasive strategy for routine clinical investigation on CTCs.

Circulating Tumor Cells in Gastrointestinal Cancers: Current Status and Future Perspectives

“…Moreover, these technologies have provided improved in situ platforms for molecular analysis by fluorescent in situ hybridization (FISH) or immunofluorescence (IF) (45), as well as for the extraction of biomolecules for downstream genomic and transcriptomic sequencing (43). In addition, these platforms also provide the opportunity for CTCs release and ex vivo expansion, which lays an important foundation to further understand the biological characteristics of CTCs (46).…”

Circulating tumor microemboli: Progress in molecular understanding and enrichment technologies

“…Chen et al introduced a novel approach for easy cell/cluster release by coating the microfluid channel surface with "supported lipid bilayer" (SLB). Coating of the device with anti-EpCAM conjugated SLB supposedly provides "non-fouling" surface which reduces non-specific interactions between cells and the channel walls thus minimizing the shear force required to detach captured cells (Chen et al, 2016). Furthermore, interactions of SLB hydrophilic lipid molecule head groups with glass surface can be easily disrupted by introduction of air bubbles thus enabling easy release of CTCs/CTM without dissociating cell-antibody binding.…”

“…The device operation consists of three steps; capture of CTCs by flowing 2 ml of blood through the microchannel at a rate of 1.5 mlh -1 ( Figure 4E, left), in second step washing with buffer at increased flow rate is used to remove cells bound non-specifically to surface, and finally disruption of SLB assembly by gentle air sweep to release viable CTCs/CTM (Chen et al, 2016). Gentle air sweep release strategy ( Figure 4E, right) not only proved to be very efficient (99.7% in 3 repeats), 86% of the released cells were viable.…”

“…Evidence has started to accumulate suggesting that the contribution of CTM to metastatic cancer spread may be far greater than previously appreciated (Aceto et al, 2014;Au et al, 2016;Cheung et al, 2016). CTM have been detected in several cancers such as; prostate (Aceto et al, 2014;Brandt et al, 1996), pancreatic (Xu et al, 2017), breast (Ozkumur et al, 2013), colorectal (Chen et al, 2016), as well as small and non-small cell lung cancers (Hou et al, 2011;Hou et al, 2012). Moreover, studies have demonstrated that presence of even a single CTM in patient blood is significantly correlated with reduced progression free survival rates (Au et al, 2017;Hou et al, 2012;Jansson et al, 2016;Mu et al, 2015).…”

Improving sensitivity and specificity of capturing and detecting targeted cancer cells with anti-biofouling polymer coated magnetic iron oxide nanoparticles

“…9–18 While conventional immuno-magnetic separation methods with micron-sized magnetic beads, such as Dynabeads ® , work well in separating and sorting cells in large number, 19,20 their efficacy in detecting and capturing the extremely small number of CTCs from samples containing a large excess of other biomolecules or non-targeted cells in the background, e.g., tens of CTCs with millions of other cells in routinely collected blood samples, remains problematic. 9,21,22 Hence, substantial efforts have been made to develop and apply magnetic nanoparticles to immuno-magnetic separation, given the advantages of nanoparticles compared with micron-sized particles, including better suspendability in the sample or media, faster cell surface receptor binding via conjugated targeting ligands, and high affinity of cell binding with more particles bound on each cell and an increased number of cell-nanoparticle interactions and therefore a higher particle-to-/cell ratio.…”