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Separable bilayer microfiltration device for viable label-free enrichment of circulating tumour cells.

Zhou MD, Hao S, Williams AJ, Harouaka RA, Schrand B, Rawal S, Ao Z, Brenneman R, Gilboa E, Lu B, Wang S, Zhu J, Datar R, Cote R, Tai YC, Zheng SY - Sci Rep (2014)

Bottom Line: Addressing this challenge, we present a separable bilayer (SB) microfilter for viable size-based CTC capture.Unlike other single-layer CTC microfilters, the precise gap between the two layers and the architecture of pore alignment result in drastic reduction in mechanical stress on CTCs, capturing them viably.In a metastatic mouse model, SB microfilters successfully enriched viable mouse CTCs from 0.4-0.6 mL whole mouse blood samples and established in vitro cultures for further genetic and functional analysis.

View Article: PubMed Central - PubMed

Affiliation: Micro &Nano Integrated Biosystem (MINIBio) Laboratory, Department of Biomedical Engineering and Materials Research Institute, Pennsylvania State University, University Park, PA 16802, U.S.A.

ABSTRACT
The analysis of circulating tumour cells (CTCs) in cancer patients could provide important information for therapeutic management. Enrichment of viable CTCs could permit performance of functional analyses on CTCs to broaden understanding of metastatic disease. However, this has not been widely accomplished. Addressing this challenge, we present a separable bilayer (SB) microfilter for viable size-based CTC capture. Unlike other single-layer CTC microfilters, the precise gap between the two layers and the architecture of pore alignment result in drastic reduction in mechanical stress on CTCs, capturing them viably. Using multiple cancer cell lines spiked in healthy donor blood, the SB microfilter demonstrated high capture efficiency (78-83%), high retention of cell viability (71-74%), high tumour cell enrichment against leukocytes (1.7-2 × 10(3)), and widespread ability to establish cultures post-capture (100% of cell lines tested). In a metastatic mouse model, SB microfilters successfully enriched viable mouse CTCs from 0.4-0.6 mL whole mouse blood samples and established in vitro cultures for further genetic and functional analysis. Our preliminary studies reflect the efficacy of the SB microfilter device to efficiently and reliably enrich viable CTCs in animal model studies, constituting an exciting technology for new insights in cancer research.

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Related in: MedlinePlus

Device design.(A): Each device has four large filter patches; (B): Each filter patch is an 8 by 8 array of elemental units; (C): Geometry of four elemental units and their arrangement. A picture of an elemental unit (boxed in orange dash lines) is shown in Figure 1C.
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f6: Device design.(A): Each device has four large filter patches; (B): Each filter patch is an 8 by 8 array of elemental units; (C): Geometry of four elemental units and their arrangement. A picture of an elemental unit (boxed in orange dash lines) is shown in Figure 1C.

Mentions: A device is composed of a 2 by 2 array of filter patches (Figure 6A). Each patch is an 8 by 8 array of the elemental units (Figure 6B). Each elemental unit is 530 μm by 530 μm in dimension. An element unit is separated from its neighbours by 50 μm (Figure 6C). These 50 μm borders are where the top and bottom parylene-C layers adhere to each other. The gap between the top and bottom parylene-C membranes can be precisely defined (5.5 μm in the current device) by the thickness of the sacrificial photoresist. The bottom layer contains the 8 μm diameter holes arranged hexagonally. The centre-to-centre distance of the 8 μm diameter holes is 25 μm. Larger holes of 40 μm diameter are created on the top layer and aligned to the centres of the corresponding hexagon patterns on the bottom layer.


Separable bilayer microfiltration device for viable label-free enrichment of circulating tumour cells.

Zhou MD, Hao S, Williams AJ, Harouaka RA, Schrand B, Rawal S, Ao Z, Brenneman R, Gilboa E, Lu B, Wang S, Zhu J, Datar R, Cote R, Tai YC, Zheng SY - Sci Rep (2014)

Device design.(A): Each device has four large filter patches; (B): Each filter patch is an 8 by 8 array of elemental units; (C): Geometry of four elemental units and their arrangement. A picture of an elemental unit (boxed in orange dash lines) is shown in Figure 1C.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260227&req=5

f6: Device design.(A): Each device has four large filter patches; (B): Each filter patch is an 8 by 8 array of elemental units; (C): Geometry of four elemental units and their arrangement. A picture of an elemental unit (boxed in orange dash lines) is shown in Figure 1C.
Mentions: A device is composed of a 2 by 2 array of filter patches (Figure 6A). Each patch is an 8 by 8 array of the elemental units (Figure 6B). Each elemental unit is 530 μm by 530 μm in dimension. An element unit is separated from its neighbours by 50 μm (Figure 6C). These 50 μm borders are where the top and bottom parylene-C layers adhere to each other. The gap between the top and bottom parylene-C membranes can be precisely defined (5.5 μm in the current device) by the thickness of the sacrificial photoresist. The bottom layer contains the 8 μm diameter holes arranged hexagonally. The centre-to-centre distance of the 8 μm diameter holes is 25 μm. Larger holes of 40 μm diameter are created on the top layer and aligned to the centres of the corresponding hexagon patterns on the bottom layer.

Bottom Line: Addressing this challenge, we present a separable bilayer (SB) microfilter for viable size-based CTC capture.Unlike other single-layer CTC microfilters, the precise gap between the two layers and the architecture of pore alignment result in drastic reduction in mechanical stress on CTCs, capturing them viably.In a metastatic mouse model, SB microfilters successfully enriched viable mouse CTCs from 0.4-0.6 mL whole mouse blood samples and established in vitro cultures for further genetic and functional analysis.

View Article: PubMed Central - PubMed

Affiliation: Micro &Nano Integrated Biosystem (MINIBio) Laboratory, Department of Biomedical Engineering and Materials Research Institute, Pennsylvania State University, University Park, PA 16802, U.S.A.

ABSTRACT
The analysis of circulating tumour cells (CTCs) in cancer patients could provide important information for therapeutic management. Enrichment of viable CTCs could permit performance of functional analyses on CTCs to broaden understanding of metastatic disease. However, this has not been widely accomplished. Addressing this challenge, we present a separable bilayer (SB) microfilter for viable size-based CTC capture. Unlike other single-layer CTC microfilters, the precise gap between the two layers and the architecture of pore alignment result in drastic reduction in mechanical stress on CTCs, capturing them viably. Using multiple cancer cell lines spiked in healthy donor blood, the SB microfilter demonstrated high capture efficiency (78-83%), high retention of cell viability (71-74%), high tumour cell enrichment against leukocytes (1.7-2 × 10(3)), and widespread ability to establish cultures post-capture (100% of cell lines tested). In a metastatic mouse model, SB microfilters successfully enriched viable mouse CTCs from 0.4-0.6 mL whole mouse blood samples and established in vitro cultures for further genetic and functional analysis. Our preliminary studies reflect the efficacy of the SB microfilter device to efficiently and reliably enrich viable CTCs in animal model studies, constituting an exciting technology for new insights in cancer research.

Show MeSH
Related in: MedlinePlus