Quantitative measurement and control of oxygen levels in microfluidic poly(dimethylsiloxane) bioreactors during cell culture

Geeta Mehta, Khamir Mehta, Dhruv Sud, Jonathan W. Song, Tommaso Bersano-Begey, Nobuyuki Futai, Yun Seok Heo, Mary Ann Mycek, Jennifer J. Linderman, Shuichi Takayama

Research output: Contribution to journalArticle

163 Citations (Scopus)

Abstract

Microfluidic bioreactors fabricated from highly gas-permeable poly(dimethylsiloxane) (PDMS) materials have been observed, somewhat unexpectedly, to give rise to heterogeneous long term responses along the length of a perfused mammalian cell culture channel, reminiscent of physiologic tissue zonation that arises at least in part due to oxygen gradients. To develop a more quantitative understanding and enable better control of the physical-chemical mechanisms underlying cell biological events in such PDMS reactors, dissolved oxygen concentrations in the channel system were quantified in real time using fluorescence intensity and lifetime imaging of an oxygen sensitive dye, ruthenium tris(2,2'-dipyridyl) dichloride hexahydrate (RTDP). The data indicate that despite oxygen diffusion through PDMS, uptake of oxygen by cells inside the perfused PDMS microchannels induces an axial oxygen concentration gradient, with lower levels recorded in downstream regions. The oxygen concentration gradient generated by a balance of cellular uptake, convective transport by media flow, and permeation through PDMS in our devices ranged from 0.0003 (mg/l)/mm to 0.7 (mg/l)/mm. The existence of such steep gradients induced by cellular uptake can have important biological consequences. Results are consistent with our mathematical model and give insight into the conditions under which flux of oxygen through PDMS into the microchannels will or will not contribute significantly to oxygen delivery to cells and also provide a design tool to manipulate and control oxygen for cell culture and device engineering. The combination of computerized microfluidics, in situ oxygen sensing, and mathematical models opens new windows for microphysiologic studies utilizing oxygen gradients and low oxygen tensions.

Original languageEnglish
Pages (from-to)123-134
Number of pages12
JournalBiomedical Microdevices
Volume9
Issue number2
DOIs
Publication statusPublished - 2007 Apr
Externally publishedYes

Fingerprint

Polydimethylsiloxane
Bioreactors
Cell culture
Microfluidics
Oxygen
Microchannels
baysilon
Mathematical models
2,2'-Dipyridyl
Ruthenium
Dissolved oxygen
Permeation
Coloring Agents
Dyes
Gases
Fluorescence

Keywords

  • Fluorescence intensity
  • Fluorescence lifetime imaging microscopy (FLIM)
  • Microbioreactor
  • Microfluidics
  • Myoblast
  • Oxygen gradients
  • Oxygen sensing
  • PDMS
  • Rutheniumtris(2,2'-dipyridyl) dichloride hexahydrate (RTDP)

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Genetics
  • Neuroscience(all)
  • Bioengineering
  • Biomedical Engineering

Cite this

Quantitative measurement and control of oxygen levels in microfluidic poly(dimethylsiloxane) bioreactors during cell culture. / Mehta, Geeta; Mehta, Khamir; Sud, Dhruv; Song, Jonathan W.; Bersano-Begey, Tommaso; Futai, Nobuyuki; Heo, Yun Seok; Mycek, Mary Ann; Linderman, Jennifer J.; Takayama, Shuichi.

In: Biomedical Microdevices, Vol. 9, No. 2, 04.2007, p. 123-134.

Research output: Contribution to journalArticle

Mehta, G, Mehta, K, Sud, D, Song, JW, Bersano-Begey, T, Futai, N, Heo, YS, Mycek, MA, Linderman, JJ & Takayama, S 2007, 'Quantitative measurement and control of oxygen levels in microfluidic poly(dimethylsiloxane) bioreactors during cell culture', Biomedical Microdevices, vol. 9, no. 2, pp. 123-134. https://doi.org/10.1007/s10544-006-9005-7
Mehta, Geeta ; Mehta, Khamir ; Sud, Dhruv ; Song, Jonathan W. ; Bersano-Begey, Tommaso ; Futai, Nobuyuki ; Heo, Yun Seok ; Mycek, Mary Ann ; Linderman, Jennifer J. ; Takayama, Shuichi. / Quantitative measurement and control of oxygen levels in microfluidic poly(dimethylsiloxane) bioreactors during cell culture. In: Biomedical Microdevices. 2007 ; Vol. 9, No. 2. pp. 123-134.
@article{4915cb7c196d43bfb0b6cd889325d5f5,
title = "Quantitative measurement and control of oxygen levels in microfluidic poly(dimethylsiloxane) bioreactors during cell culture",
abstract = "Microfluidic bioreactors fabricated from highly gas-permeable poly(dimethylsiloxane) (PDMS) materials have been observed, somewhat unexpectedly, to give rise to heterogeneous long term responses along the length of a perfused mammalian cell culture channel, reminiscent of physiologic tissue zonation that arises at least in part due to oxygen gradients. To develop a more quantitative understanding and enable better control of the physical-chemical mechanisms underlying cell biological events in such PDMS reactors, dissolved oxygen concentrations in the channel system were quantified in real time using fluorescence intensity and lifetime imaging of an oxygen sensitive dye, ruthenium tris(2,2'-dipyridyl) dichloride hexahydrate (RTDP). The data indicate that despite oxygen diffusion through PDMS, uptake of oxygen by cells inside the perfused PDMS microchannels induces an axial oxygen concentration gradient, with lower levels recorded in downstream regions. The oxygen concentration gradient generated by a balance of cellular uptake, convective transport by media flow, and permeation through PDMS in our devices ranged from 0.0003 (mg/l)/mm to 0.7 (mg/l)/mm. The existence of such steep gradients induced by cellular uptake can have important biological consequences. Results are consistent with our mathematical model and give insight into the conditions under which flux of oxygen through PDMS into the microchannels will or will not contribute significantly to oxygen delivery to cells and also provide a design tool to manipulate and control oxygen for cell culture and device engineering. The combination of computerized microfluidics, in situ oxygen sensing, and mathematical models opens new windows for microphysiologic studies utilizing oxygen gradients and low oxygen tensions.",
keywords = "Fluorescence intensity, Fluorescence lifetime imaging microscopy (FLIM), Microbioreactor, Microfluidics, Myoblast, Oxygen gradients, Oxygen sensing, PDMS, Rutheniumtris(2,2'-dipyridyl) dichloride hexahydrate (RTDP)",
author = "Geeta Mehta and Khamir Mehta and Dhruv Sud and Song, {Jonathan W.} and Tommaso Bersano-Begey and Nobuyuki Futai and Heo, {Yun Seok} and Mycek, {Mary Ann} and Linderman, {Jennifer J.} and Shuichi Takayama",
year = "2007",
month = "4",
doi = "10.1007/s10544-006-9005-7",
language = "English",
volume = "9",
pages = "123--134",
journal = "Biomedical Microdevices",
issn = "1387-2176",
publisher = "Kluwer Academic Publishers",
number = "2",

}

TY - JOUR

T1 - Quantitative measurement and control of oxygen levels in microfluidic poly(dimethylsiloxane) bioreactors during cell culture

AU - Mehta, Geeta

AU - Mehta, Khamir

AU - Sud, Dhruv

AU - Song, Jonathan W.

AU - Bersano-Begey, Tommaso

AU - Futai, Nobuyuki

AU - Heo, Yun Seok

AU - Mycek, Mary Ann

AU - Linderman, Jennifer J.

AU - Takayama, Shuichi

PY - 2007/4

Y1 - 2007/4

N2 - Microfluidic bioreactors fabricated from highly gas-permeable poly(dimethylsiloxane) (PDMS) materials have been observed, somewhat unexpectedly, to give rise to heterogeneous long term responses along the length of a perfused mammalian cell culture channel, reminiscent of physiologic tissue zonation that arises at least in part due to oxygen gradients. To develop a more quantitative understanding and enable better control of the physical-chemical mechanisms underlying cell biological events in such PDMS reactors, dissolved oxygen concentrations in the channel system were quantified in real time using fluorescence intensity and lifetime imaging of an oxygen sensitive dye, ruthenium tris(2,2'-dipyridyl) dichloride hexahydrate (RTDP). The data indicate that despite oxygen diffusion through PDMS, uptake of oxygen by cells inside the perfused PDMS microchannels induces an axial oxygen concentration gradient, with lower levels recorded in downstream regions. The oxygen concentration gradient generated by a balance of cellular uptake, convective transport by media flow, and permeation through PDMS in our devices ranged from 0.0003 (mg/l)/mm to 0.7 (mg/l)/mm. The existence of such steep gradients induced by cellular uptake can have important biological consequences. Results are consistent with our mathematical model and give insight into the conditions under which flux of oxygen through PDMS into the microchannels will or will not contribute significantly to oxygen delivery to cells and also provide a design tool to manipulate and control oxygen for cell culture and device engineering. The combination of computerized microfluidics, in situ oxygen sensing, and mathematical models opens new windows for microphysiologic studies utilizing oxygen gradients and low oxygen tensions.

AB - Microfluidic bioreactors fabricated from highly gas-permeable poly(dimethylsiloxane) (PDMS) materials have been observed, somewhat unexpectedly, to give rise to heterogeneous long term responses along the length of a perfused mammalian cell culture channel, reminiscent of physiologic tissue zonation that arises at least in part due to oxygen gradients. To develop a more quantitative understanding and enable better control of the physical-chemical mechanisms underlying cell biological events in such PDMS reactors, dissolved oxygen concentrations in the channel system were quantified in real time using fluorescence intensity and lifetime imaging of an oxygen sensitive dye, ruthenium tris(2,2'-dipyridyl) dichloride hexahydrate (RTDP). The data indicate that despite oxygen diffusion through PDMS, uptake of oxygen by cells inside the perfused PDMS microchannels induces an axial oxygen concentration gradient, with lower levels recorded in downstream regions. The oxygen concentration gradient generated by a balance of cellular uptake, convective transport by media flow, and permeation through PDMS in our devices ranged from 0.0003 (mg/l)/mm to 0.7 (mg/l)/mm. The existence of such steep gradients induced by cellular uptake can have important biological consequences. Results are consistent with our mathematical model and give insight into the conditions under which flux of oxygen through PDMS into the microchannels will or will not contribute significantly to oxygen delivery to cells and also provide a design tool to manipulate and control oxygen for cell culture and device engineering. The combination of computerized microfluidics, in situ oxygen sensing, and mathematical models opens new windows for microphysiologic studies utilizing oxygen gradients and low oxygen tensions.

KW - Fluorescence intensity

KW - Fluorescence lifetime imaging microscopy (FLIM)

KW - Microbioreactor

KW - Microfluidics

KW - Myoblast

KW - Oxygen gradients

KW - Oxygen sensing

KW - PDMS

KW - Rutheniumtris(2,2'-dipyridyl) dichloride hexahydrate (RTDP)

UR - http://www.scopus.com/inward/record.url?scp=34147115167&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34147115167&partnerID=8YFLogxK

U2 - 10.1007/s10544-006-9005-7

DO - 10.1007/s10544-006-9005-7

M3 - Article

C2 - 17160707

AN - SCOPUS:34147115167

VL - 9

SP - 123

EP - 134

JO - Biomedical Microdevices

JF - Biomedical Microdevices

SN - 1387-2176

IS - 2

ER -