Abstract

Regions of hypoxia are common in solid tumors and are associated with enhanced malignancy, metastasis, and chemo/radio resistance. Real-time hypoxic cellular experimentation is challenging due to the constant need for oxygen control. Most microfluidic platforms developed thus far for hypoxic cell studies are burdened by complex design parameters and are difficult to use for uninitiated investigators. However, open-well microfluidic platforms enable short and long term hypoxic cell studies with an ease of use workflow. Specifically, open-well platforms enable manipulation and addition of cells, media, and reagents using a micropipette for hypoxic cell studies in tunable dissolved oxygen concentrations as low 0.3mg/l. We analyzed design considerations for open-well microfluidic platforms such as media height, membrane thickness, and impermeable barriers to determine their effects on the amount of dissolved oxygen within the platform. The oxygen concentration was determined by experimental measurements and computational simulations. To examine cell behavior under controlled oxygen conditions, hypoxia-induced changes to hypoxia inducible factor activity and the mitochondrial redox environment were studied. A fluorescent reporter construct was used to monitor the stabilization of hypoxia inducible factors 1α and 2α throughout chronic hypoxia. Reporter construct fluorescence intensity inversely correlated with dissolved oxygen in the medium, as expected. Additionally, the glutathione redox poise of the mitochondrial matrix in living cancer cells was monitored throughout acute hypoxia with a genetically encoded redox probe and was observed to undergo a reductive response to hypoxia. Overall, these studies validate an easy to use open-well platform suitable for studying complex cell behaviors in hypoxia.

Original languageEnglish (US)
Article number054116
JournalBiomicrofluidics
Volume11
Issue number5
DOIs
StatePublished - Sep 1 2017

Fingerprint

Microfluidics
hypoxia
Dissolved oxygen
platforms
Cells
Oxygen
oxygen
Oxidation-Reduction
Hypoxia-Inducible Factor 1
Glutathione
Tumors
Stabilization
cells
Fluorescence
Neoplasms
Membranes
Workflow
glutathione
Hypoxia
Radio

ASJC Scopus subject areas

  • Molecular Biology
  • Materials Science(all)
  • Genetics
  • Condensed Matter Physics
  • Physical and Theoretical Chemistry

Cite this

Design considerations for open-well microfluidic platforms for hypoxic cell studies. / Byrne, Matthew B.; Leslie, Matthew T.; Patel, Heeral S.; Gaskins, H Rex; Kenis, Paul J A.

In: Biomicrofluidics, Vol. 11, No. 5, 054116, 01.09.2017.

Research output: Contribution to journalArticle

Byrne, Matthew B. ; Leslie, Matthew T. ; Patel, Heeral S. ; Gaskins, H Rex ; Kenis, Paul J A. / Design considerations for open-well microfluidic platforms for hypoxic cell studies. In: Biomicrofluidics. 2017 ; Vol. 11, No. 5.
@article{02b581c91123489e9bf468d13335354c,
title = "Design considerations for open-well microfluidic platforms for hypoxic cell studies",
abstract = "Regions of hypoxia are common in solid tumors and are associated with enhanced malignancy, metastasis, and chemo/radio resistance. Real-time hypoxic cellular experimentation is challenging due to the constant need for oxygen control. Most microfluidic platforms developed thus far for hypoxic cell studies are burdened by complex design parameters and are difficult to use for uninitiated investigators. However, open-well microfluidic platforms enable short and long term hypoxic cell studies with an ease of use workflow. Specifically, open-well platforms enable manipulation and addition of cells, media, and reagents using a micropipette for hypoxic cell studies in tunable dissolved oxygen concentrations as low 0.3mg/l. We analyzed design considerations for open-well microfluidic platforms such as media height, membrane thickness, and impermeable barriers to determine their effects on the amount of dissolved oxygen within the platform. The oxygen concentration was determined by experimental measurements and computational simulations. To examine cell behavior under controlled oxygen conditions, hypoxia-induced changes to hypoxia inducible factor activity and the mitochondrial redox environment were studied. A fluorescent reporter construct was used to monitor the stabilization of hypoxia inducible factors 1α and 2α throughout chronic hypoxia. Reporter construct fluorescence intensity inversely correlated with dissolved oxygen in the medium, as expected. Additionally, the glutathione redox poise of the mitochondrial matrix in living cancer cells was monitored throughout acute hypoxia with a genetically encoded redox probe and was observed to undergo a reductive response to hypoxia. Overall, these studies validate an easy to use open-well platform suitable for studying complex cell behaviors in hypoxia.",
author = "Byrne, {Matthew B.} and Leslie, {Matthew T.} and Patel, {Heeral S.} and Gaskins, {H Rex} and Kenis, {Paul J A}",
year = "2017",
month = "9",
day = "1",
doi = "10.1063/1.4998579",
language = "English (US)",
volume = "11",
journal = "Biomicrofluidics",
issn = "1932-1058",
publisher = "American Institute of Physics Publising LLC",
number = "5",

}

TY - JOUR

T1 - Design considerations for open-well microfluidic platforms for hypoxic cell studies

AU - Byrne, Matthew B.

AU - Leslie, Matthew T.

AU - Patel, Heeral S.

AU - Gaskins, H Rex

AU - Kenis, Paul J A

PY - 2017/9/1

Y1 - 2017/9/1

N2 - Regions of hypoxia are common in solid tumors and are associated with enhanced malignancy, metastasis, and chemo/radio resistance. Real-time hypoxic cellular experimentation is challenging due to the constant need for oxygen control. Most microfluidic platforms developed thus far for hypoxic cell studies are burdened by complex design parameters and are difficult to use for uninitiated investigators. However, open-well microfluidic platforms enable short and long term hypoxic cell studies with an ease of use workflow. Specifically, open-well platforms enable manipulation and addition of cells, media, and reagents using a micropipette for hypoxic cell studies in tunable dissolved oxygen concentrations as low 0.3mg/l. We analyzed design considerations for open-well microfluidic platforms such as media height, membrane thickness, and impermeable barriers to determine their effects on the amount of dissolved oxygen within the platform. The oxygen concentration was determined by experimental measurements and computational simulations. To examine cell behavior under controlled oxygen conditions, hypoxia-induced changes to hypoxia inducible factor activity and the mitochondrial redox environment were studied. A fluorescent reporter construct was used to monitor the stabilization of hypoxia inducible factors 1α and 2α throughout chronic hypoxia. Reporter construct fluorescence intensity inversely correlated with dissolved oxygen in the medium, as expected. Additionally, the glutathione redox poise of the mitochondrial matrix in living cancer cells was monitored throughout acute hypoxia with a genetically encoded redox probe and was observed to undergo a reductive response to hypoxia. Overall, these studies validate an easy to use open-well platform suitable for studying complex cell behaviors in hypoxia.

AB - Regions of hypoxia are common in solid tumors and are associated with enhanced malignancy, metastasis, and chemo/radio resistance. Real-time hypoxic cellular experimentation is challenging due to the constant need for oxygen control. Most microfluidic platforms developed thus far for hypoxic cell studies are burdened by complex design parameters and are difficult to use for uninitiated investigators. However, open-well microfluidic platforms enable short and long term hypoxic cell studies with an ease of use workflow. Specifically, open-well platforms enable manipulation and addition of cells, media, and reagents using a micropipette for hypoxic cell studies in tunable dissolved oxygen concentrations as low 0.3mg/l. We analyzed design considerations for open-well microfluidic platforms such as media height, membrane thickness, and impermeable barriers to determine their effects on the amount of dissolved oxygen within the platform. The oxygen concentration was determined by experimental measurements and computational simulations. To examine cell behavior under controlled oxygen conditions, hypoxia-induced changes to hypoxia inducible factor activity and the mitochondrial redox environment were studied. A fluorescent reporter construct was used to monitor the stabilization of hypoxia inducible factors 1α and 2α throughout chronic hypoxia. Reporter construct fluorescence intensity inversely correlated with dissolved oxygen in the medium, as expected. Additionally, the glutathione redox poise of the mitochondrial matrix in living cancer cells was monitored throughout acute hypoxia with a genetically encoded redox probe and was observed to undergo a reductive response to hypoxia. Overall, these studies validate an easy to use open-well platform suitable for studying complex cell behaviors in hypoxia.

UR - http://www.scopus.com/inward/record.url?scp=85032740423&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85032740423&partnerID=8YFLogxK

U2 - 10.1063/1.4998579

DO - 10.1063/1.4998579

M3 - Article

AN - SCOPUS:85032740423

VL - 11

JO - Biomicrofluidics

JF - Biomicrofluidics

SN - 1932-1058

IS - 5

M1 - 054116

ER -