Limits...
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.

Show MeSH

Related in: MedlinePlus

Device design.(A): Cartoon of device cross-sectional view showing tumour cells are captured along the edges of the large top parylene-C pores; (B): 3D view of an elemental unit model with key geometrical parameters labelled, including the gap distance in inset; (C): Microscopic picture of top view of an elemental unit showing large pores on top parylene-C layer and small pores on bottom parylene-C layer with index dots on the top left corner. Scale bar is 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4260227&req=5

f1: Device design.(A): Cartoon of device cross-sectional view showing tumour cells are captured along the edges of the large top parylene-C pores; (B): 3D view of an elemental unit model with key geometrical parameters labelled, including the gap distance in inset; (C): Microscopic picture of top view of an elemental unit showing large pores on top parylene-C layer and small pores on bottom parylene-C layer with index dots on the top left corner. Scale bar is 100 μm.

Mentions: In the SB microfilter, the capture is realized by the gap between the top and bottom porous membranes (Figure 1A). They were made from biocompatible polymer parylene-C aiming to better preserve the viability of the cells. A 3D view in Figure 1B summarizes the geometrical design parameters. The bottom layer contains the 8 μm diameter holes arranged hexagonally. Larger holes of 40 μm diameter were created on the top parylene-C layer and aligned to the centres of the corresponding hexagon patterns on the bottom layer. An example elemental unit after fabrication is shown in Figure 1C. Each elemental unit is encoded by dots at the top left corner of the unit on the bottom parylene-C layer as an index to facilitate the identification and positioning of captured cells of interest.


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): Cartoon of device cross-sectional view showing tumour cells are captured along the edges of the large top parylene-C pores; (B): 3D view of an elemental unit model with key geometrical parameters labelled, including the gap distance in inset; (C): Microscopic picture of top view of an elemental unit showing large pores on top parylene-C layer and small pores on bottom parylene-C layer with index dots on the top left corner. Scale bar is 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Device design.(A): Cartoon of device cross-sectional view showing tumour cells are captured along the edges of the large top parylene-C pores; (B): 3D view of an elemental unit model with key geometrical parameters labelled, including the gap distance in inset; (C): Microscopic picture of top view of an elemental unit showing large pores on top parylene-C layer and small pores on bottom parylene-C layer with index dots on the top left corner. Scale bar is 100 μm.
Mentions: In the SB microfilter, the capture is realized by the gap between the top and bottom porous membranes (Figure 1A). They were made from biocompatible polymer parylene-C aiming to better preserve the viability of the cells. A 3D view in Figure 1B summarizes the geometrical design parameters. The bottom layer contains the 8 μm diameter holes arranged hexagonally. Larger holes of 40 μm diameter were created on the top parylene-C layer and aligned to the centres of the corresponding hexagon patterns on the bottom layer. An example elemental unit after fabrication is shown in Figure 1C. Each elemental unit is encoded by dots at the top left corner of the unit on the bottom parylene-C layer as an index to facilitate the identification and positioning of captured cells of interest.

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