Single-cell proteolytic activity measurement using microfluidics for rare cell populations

Yu Chih Chen; Euisik Yoon

doi:10.1016/bs.mie.2019.07.012

Single-cell proteolytic activity measurement using microfluidics for rare cell populations

Yu Chih Chen, Euisik Yoon

Yonsei Advanced Science Institute

Research output: Chapter in Book/Report/Conference proceeding › Chapter

Abstract

Proteolytic degradation of the extracellular matrix represents a key step in cancer dissemination and metastasis. To probe cellular proteolytic activity, fluorescent sensing substrate was developed, yet prior studies focused on average activity of thousands of cells. Considerable evidence suggests a specialized subset of cancer cells are driving metastasis, highlighting the value of single-cell approach to reveal cancer cellular heterogeneity. In addition, when only a small number of cells are available, single-cell analysis is required to draw a statistical conclusion. Here, we present a microfluidic platform that provides high-efficiency cell loading and simple valveless isolation, so the proteolytic activity of a small number (10–100) of cells can be individually characterized. Furthermore, the platform allows monitoring single cells at multiple time points for the investigation of dynamics in proteolytic activity. The presented platform represents a simple and reliable tool for single-cell proteolytic analysis, illuminating the heterogeneous and dynamic nature of cancer cells.

Original language	English
Title of host publication	Enzyme Activity in Single Cells
Editors	Nancy L. Allbritton, Michelle L. Kovarik
Publisher	Academic Press Inc.
Pages	129-143
Number of pages	15
ISBN (Print)	9780128170908
DOIs	https://doi.org/10.1016/bs.mie.2019.07.012
Publication status	Published - 2019

Publication series

Name	Methods in Enzymology
Volume	628
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Bibliographical note

Funding Information:
This work was supported by the grants R01 CA 203810 and R21 CA 195016 from NIH to E.Y. The support of Y.-C.C. from Forbes Institute for Cancer Discovery is also acknowledged. We thank the Lurie Nanofabrication Facility of the University of Michigan (Ann Arbor, MI) for device fabrication.

Funding Information:
This work was supported by the grants R01 CA 203810 and R21 CA 195016 from NIH to E.Y. The support of Y.-C.C. from Forbes Institute for Cancer Discovery is also acknowledged. We thank the Lurie Nanofabrication Facility of the University of Michigan (Ann Arbor, MI) for device fabrication.

Publisher Copyright:
© 2019 Elsevier Inc.

All Science Journal Classification (ASJC) codes

Biochemistry
Molecular Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/bs.mie.2019.07.012

Cite this

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title = "Single-cell proteolytic activity measurement using microfluidics for rare cell populations",

abstract = "Proteolytic degradation of the extracellular matrix represents a key step in cancer dissemination and metastasis. To probe cellular proteolytic activity, fluorescent sensing substrate was developed, yet prior studies focused on average activity of thousands of cells. Considerable evidence suggests a specialized subset of cancer cells are driving metastasis, highlighting the value of single-cell approach to reveal cancer cellular heterogeneity. In addition, when only a small number of cells are available, single-cell analysis is required to draw a statistical conclusion. Here, we present a microfluidic platform that provides high-efficiency cell loading and simple valveless isolation, so the proteolytic activity of a small number (10–100) of cells can be individually characterized. Furthermore, the platform allows monitoring single cells at multiple time points for the investigation of dynamics in proteolytic activity. The presented platform represents a simple and reliable tool for single-cell proteolytic analysis, illuminating the heterogeneous and dynamic nature of cancer cells.",

author = "Chen, {Yu Chih} and Euisik Yoon",

note = "Funding Information: This work was supported by the grants R01 CA 203810 and R21 CA 195016 from NIH to E.Y. The support of Y.-C.C. from Forbes Institute for Cancer Discovery is also acknowledged. We thank the Lurie Nanofabrication Facility of the University of Michigan (Ann Arbor, MI) for device fabrication. Funding Information: This work was supported by the grants R01 CA 203810 and R21 CA 195016 from NIH to E.Y. The support of Y.-C.C. from Forbes Institute for Cancer Discovery is also acknowledged. We thank the Lurie Nanofabrication Facility of the University of Michigan (Ann Arbor, MI) for device fabrication. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",

year = "2019",

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language = "English",

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N2 - Proteolytic degradation of the extracellular matrix represents a key step in cancer dissemination and metastasis. To probe cellular proteolytic activity, fluorescent sensing substrate was developed, yet prior studies focused on average activity of thousands of cells. Considerable evidence suggests a specialized subset of cancer cells are driving metastasis, highlighting the value of single-cell approach to reveal cancer cellular heterogeneity. In addition, when only a small number of cells are available, single-cell analysis is required to draw a statistical conclusion. Here, we present a microfluidic platform that provides high-efficiency cell loading and simple valveless isolation, so the proteolytic activity of a small number (10–100) of cells can be individually characterized. Furthermore, the platform allows monitoring single cells at multiple time points for the investigation of dynamics in proteolytic activity. The presented platform represents a simple and reliable tool for single-cell proteolytic analysis, illuminating the heterogeneous and dynamic nature of cancer cells.

AB - Proteolytic degradation of the extracellular matrix represents a key step in cancer dissemination and metastasis. To probe cellular proteolytic activity, fluorescent sensing substrate was developed, yet prior studies focused on average activity of thousands of cells. Considerable evidence suggests a specialized subset of cancer cells are driving metastasis, highlighting the value of single-cell approach to reveal cancer cellular heterogeneity. In addition, when only a small number of cells are available, single-cell analysis is required to draw a statistical conclusion. Here, we present a microfluidic platform that provides high-efficiency cell loading and simple valveless isolation, so the proteolytic activity of a small number (10–100) of cells can be individually characterized. Furthermore, the platform allows monitoring single cells at multiple time points for the investigation of dynamics in proteolytic activity. The presented platform represents a simple and reliable tool for single-cell proteolytic analysis, illuminating the heterogeneous and dynamic nature of cancer cells.

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Single-cell proteolytic activity measurement using microfluidics for rare cell populations

Abstract

Publication series

Bibliographical note

All Science Journal Classification (ASJC) codes

UN SDGs

Access to Document

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