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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

Demonstration of the operation of the SB microfilter.(A)–(B): Representative areas of the SB microfilter after filtration. MCF-7 breast cancer cells were GFP-expressing (A) while surrounding blood cells could also be seen in bright field/fluorescence composite view (B). Scale bars are 50 μm. (C)–(G): The top and bottom parylene-C membrane layers can be separated after cell enrichment. (C): Image showing the concept of separable microfilter. (D): MDA-MB-231 cells labelled with CFDA-SE. (E): Cells captured on SB microfilter before separation of the top and bottom parylene-C layers. (F): The bottom parylene-C layer after separation. (G): The top parylene-C layer after separation. Scale bars are 100 μm.
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f2: Demonstration of the operation of the SB microfilter.(A)–(B): Representative areas of the SB microfilter after filtration. MCF-7 breast cancer cells were GFP-expressing (A) while surrounding blood cells could also be seen in bright field/fluorescence composite view (B). Scale bars are 50 μm. (C)–(G): The top and bottom parylene-C membrane layers can be separated after cell enrichment. (C): Image showing the concept of separable microfilter. (D): MDA-MB-231 cells labelled with CFDA-SE. (E): Cells captured on SB microfilter before separation of the top and bottom parylene-C layers. (F): The bottom parylene-C layer after separation. (G): The top parylene-C layer after separation. Scale bars are 100 μm.

Mentions: The operation of the SB microfilter was first demonstrated using healthy blood samples spiked with tumour cell line. GFP expressing MCF-7 cells (green fluorescent) were spiked in whole blood and passed through the SB microfilter. An example of the microfilter following filtration is shown in Figure 2A&B. The majority of the captured tumour cells were located along the edges of the large top pores, although tumour cells could partially or completely wedge into the gap between the top and bottom parylene-C layers.


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)

Demonstration of the operation of the SB microfilter.(A)–(B): Representative areas of the SB microfilter after filtration. MCF-7 breast cancer cells were GFP-expressing (A) while surrounding blood cells could also be seen in bright field/fluorescence composite view (B). Scale bars are 50 μm. (C)–(G): The top and bottom parylene-C membrane layers can be separated after cell enrichment. (C): Image showing the concept of separable microfilter. (D): MDA-MB-231 cells labelled with CFDA-SE. (E): Cells captured on SB microfilter before separation of the top and bottom parylene-C layers. (F): The bottom parylene-C layer after separation. (G): The top parylene-C layer after separation. Scale bars are 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Demonstration of the operation of the SB microfilter.(A)–(B): Representative areas of the SB microfilter after filtration. MCF-7 breast cancer cells were GFP-expressing (A) while surrounding blood cells could also be seen in bright field/fluorescence composite view (B). Scale bars are 50 μm. (C)–(G): The top and bottom parylene-C membrane layers can be separated after cell enrichment. (C): Image showing the concept of separable microfilter. (D): MDA-MB-231 cells labelled with CFDA-SE. (E): Cells captured on SB microfilter before separation of the top and bottom parylene-C layers. (F): The bottom parylene-C layer after separation. (G): The top parylene-C layer after separation. Scale bars are 100 μm.
Mentions: The operation of the SB microfilter was first demonstrated using healthy blood samples spiked with tumour cell line. GFP expressing MCF-7 cells (green fluorescent) were spiked in whole blood and passed through the SB microfilter. An example of the microfilter following filtration is shown in Figure 2A&B. The majority of the captured tumour cells were located along the edges of the large top pores, although tumour cells could partially or completely wedge into the gap between the top and bottom parylene-C layers.

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