Skip to main content
SpringerLink
Log in

Rapid Method for Assessing Oxygen Consumption Rate of Cells from Transient-state Measurements of Pericellular Dissolved Oxygen Concentration

  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Recently we described a method for estimating the oxygen consumption rate (OCR) of cells in static culture from equilibrium measurements of dissolved oxygen concentration (dO2), using an oxygen-sensing microplate and the steady-state solution to Fick's Law (Guarino et al. 2004). Here we describe a complementary method for estimating OCR from the transient-state rate of change of measured dO2. Although the system is open to the atmosphere and subject to a significant lag in sensor response, the rate of change of the measured dO2 immediately after seeding correlated directly with both cell number and steady-state OCR. This transient-state method is linear with cell number to a much higher density than is possible with the steady-state method because it derives from measurements made before diffusion limitations can be established. For a given sensor thickness, the same correlation line between the transient and non-diffusion-limited steady-state estimates of OCR was found to apply for various preparations of rat hepatocytes. The correlation slope varied predictably with sensor thickness. Thus, despite the non-idealities of this system, the initial rate measurement offers a rapid method to obtain an estimate of absolute OCR. To demonstrate the utility of this method, we purposefully treated rat hepatocytes in ways expected to change OCR. Cells deprived of oxygen by storage under several centimeters of medium showed decreases in both OCR and viability with time. Likewise, the OCR of hepatocytes exposed to the oxidative phosphorylation inhibitor rotenone decreased, whereas those exposed to the uncoupler dinoseb increased.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • J. Alderman J. Hynes S.M. Floyd J. Kruger R. O’Connor D.B. Papkovsky (2004) ArticleTitleA low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing Biosens. Bioelectron. 19 1529–1535 Occurrence Handle15093226 Occurrence Handle1:CAS:528:DC%2BD2cXjtFantr4%3D Occurrence Handle10.1016/j.bios.2003.12.008

    Article  PubMed  CAS  Google Scholar 

  • R.R. Deshpande E. Heinzle (2004) ArticleTitleOn-line oxygen uptake rate and culture viability measurement of animal cell culture using microplates with integrated oxygen sensors Biotechnol. Lett. 26 763–767 Occurrence Handle15195979 Occurrence Handle1:CAS:528:DC%2BD2cXjtFaksL8%3D Occurrence Handle10.1023/B:BILE.0000024101.57683.6d

    Article  PubMed  CAS  Google Scholar 

  • Fraker C., Timmins M.R., Guarino R.D., Ichii H., Molano D., Poggioli R., Pileggi A., Presnell S.C., Inverardi L. and Ricordi C. 2005. The use of the BD oxygen biosensor system to assess isolated islets of langerhans: oxygen consumption as a potential measure of islet potency. Cell Transplant. (manuscript submitted).

  • J. Genzer K. Efimenko (2000) ArticleTitleCreating long-lived superhydrophobic polymer surfaces through mechanically assembled monolayers Science 290 2130–2133 Occurrence Handle11118144 Occurrence Handle1:CAS:528:DC%2BD3cXptVyis7Y%3D Occurrence Handle10.1126/science.290.5499.2130

    Article  PubMed  CAS  Google Scholar 

  • R.D. Guarino L.E. Dike T.A. Haq J.A. Rowley J.B. Pitner M.R. Timmins (2004) ArticleTitleMethod for determining oxygen consumption rates of static cultures from microplate measurements of pericellular dissolved oxygen concentration Biotechnol. Bioeng. 86 IssueID7 775–787 Occurrence Handle15162453 Occurrence Handle1:CAS:528:DC%2BD2cXkslCgt7g%3D Occurrence Handle10.1002/bit.20072

    Article  PubMed  CAS  Google Scholar 

  • R.D. Guarino L.E. Dike T.A. Haq J.A. Rowley J.B. Pitner M.R. Timmins (2005) ArticleTitleErratumMethod for determining oxygen consumption rates of static cultures from microplate measurements of pericellular dissolved oxygen concentration Biotechnol. Bioeng. 91 IssueID3 392 Occurrence Handle1:CAS:528:DC%2BD2MXntlCnsLo%3D Occurrence Handle10.1002/bit.20613

    Article  CAS  Google Scholar 

  • D.M. Harvitt J.A. Bonanno (1998) ArticleTitleOxygen consumption of the rabbit cornea Invest. Ophthalmol. Vis. Sci. 39 IssueID2 444–448 Occurrence Handle9478006 Occurrence Handle1:STN:280:DyaK1c7jvVCqsQ%3D%3D

    PubMed  CAS  Google Scholar 

  • Dv. Heimburg K. Hemmrich S. Zachariah H. Staiger N. Pallua (2005) ArticleTitleOxygen consumption in undifferentiated versus differentiated adipogenic mesenchymal precursor cells Respir. Physiol. Neurobiol. 146 107–116 Occurrence Handle10.1016/j.resp.2004.12.013 Occurrence Handle1:CAS:528:DC%2BD2MXit1CjsLY%3D

    Article  CAS  Google Scholar 

  • F.D. Houghton J.G. Thompson C.J. Kennedy H.J. Leese (1996) ArticleTitleOxygen consumption and energy metabolism of the early mouse embryo Mol. Reprod. Dev. 44 476–485 Occurrence Handle8844690 Occurrence Handle1:CAS:528:DyaK28XkvFGrtLY%3D Occurrence Handle10.1002/(SICI)1098-2795(199608)44:4<476::AID-MRD7>3.0.CO;2-I

    Article  PubMed  CAS  Google Scholar 

  • B. Hutter G.T. John (2004) ArticleTitleEvaluation of OxoPlate for real-time assessment of antibacterial activities Curr. Microbiol. 48 57–61 Occurrence Handle15018104 Occurrence Handle1:CAS:528:DC%2BD2cXhsFKitLw%3D Occurrence Handle10.1007/s00284-003-4095-4

    Article  PubMed  CAS  Google Scholar 

  • J. Hynes S. Floyd A.E. Soini R. O’Connor D.B. Papkovsky (2003) ArticleTitleFluorescence-based cell viability screening assays using water-soluble oxygen probes J. Biomol. Screen. 8 IssueID3 264–272 Occurrence Handle12857380 Occurrence Handle1:CAS:528:DC%2BD3sXlsVegsLg%3D Occurrence Handle10.1177/1087057103008003004

    Article  PubMed  CAS  Google Scholar 

  • H.C. Isom I. Georgoff (1984) ArticleTitleQuantitative assay for albumin-producing liver cells after simian virus 40 transformation of rat hepatocytes maintained in chemically defined medium Proc. Natl. Acad. Sci. USA 81 6378–6382 Occurrence Handle6093099 Occurrence Handle1:CAS:528:DyaL2MXjvFOl Occurrence Handle10.1073/pnas.81.20.6378

    Article  PubMed  CAS  Google Scholar 

  • P. Jorjani S.S. Ozturk (1999) ArticleTitleEffects of cell density and temperature on oxygen consumption rate for different mammalian cell lines Biotechnol. Bioeng. 64 349–356 Occurrence Handle10397872 Occurrence Handle1:CAS:528:DyaK1MXktVymsbo%3D Occurrence Handle10.1002/(SICI)1097-0290(19990805)64:3<349::AID-BIT11>3.0.CO;2-V

    Article  PubMed  CAS  Google Scholar 

  • C.A. Kindig K.M. Kelley R.A. Howlett C.M. Stary M.C. Hogan (2003a) ArticleTitleAssessment of O2 uptake dynamics in isolated single skeletal myocytes J. Appl. Physiol. 94 353–357

    Google Scholar 

  • C.A. Kindig K.M. Kelley R.A. Howlett C.M. Stary M.C. Hogan (2003b) ArticleTitleAssessment of O2 uptake dynamics in isolated single skeletal myocytes J. Appl. Physiol. 94 353–357

    Google Scholar 

  • Y. Li L.S. Green R. Holtzapffel D.A. Day F.J. Bergersen (2001) ArticleTitleSupply of O2 regulates demand for O2uptake of malate by N2-fixing bacteroids from soybean nodules Microbiology 147 IssueIDPt 3 663–670 Occurrence Handle11238973 Occurrence Handle1:CAS:528:DC%2BD3MXis1OgtLY%3D

    PubMed  CAS  Google Scholar 

  • K. Mamchaoui G. Saumon (2000) ArticleTitleA method for measuring the oxygen consumption of intact cell monolayers Am. J. Physiol Lung Cell Mol. Physiol. 278 L858–863 Occurrence Handle10749764 Occurrence Handle1:CAS:528:DC%2BD3cXis12hsbs%3D

    PubMed  CAS  Google Scholar 

  • A. Matos J.L. Garland (2005) ArticleTitleEffects of community versus single strain inoculants on the biocontrol of Salmonella and microbial community dynamics in alfalfa sprouts J. Food Prot. 68 40–48 Occurrence Handle15690802

    PubMed  Google Scholar 

  • W.F. McLimans L.E. Blumenson K.V. Tunnah (1968a) ArticleTitleKinetics of gas diffusion in mammalian cell culture systems. II. Theory Biotechnol. Bioeng. 10 741–763 Occurrence Handle1:CAS:528:DyaF1MXksFCqsw%3D%3D Occurrence Handle10.1002/bit.260100604

    Article  CAS  Google Scholar 

  • W.F. McLimans E.J. Crouse K.V. Tunnah G.E. Moore (1968b) ArticleTitleKinetics of gas diffusion in mammalian cell culture systems I. Experimental Biotechnol. Bioeng. 10 725–740 Occurrence Handle1:CAS:528:DyaF1MXksFCqsg%3D%3D Occurrence Handle10.1002/bit.260100603

    Article  CAS  Google Scholar 

  • E. Metzen M. Wolff J. Fandrey W. Jelkmann (1995) ArticleTitlePericellular PO2 and O2 consumption in monolayer cell cultures Respir. Physiol. 100 101–106 Occurrence Handle7624611 Occurrence Handle1:CAS:528:DyaK2MXmt1Kqsr4%3D Occurrence Handle10.1016/0034-5687(94)00125-J

    Article  PubMed  CAS  Google Scholar 

  • T. O’Riordan D. Buckley V. Ogurtsov R. O’Connor D. Papkovsky (2000) ArticleTitleA cell viability assay based on monitoring respiration by optical oxygen sensing Anal. Biochem. 278 221–227 Occurrence Handle10660466 Occurrence Handle1:CAS:528:DC%2BD3cXoslGqtw%3D%3D Occurrence Handle10.1006/abio.1999.4431

    Article  PubMed  CAS  Google Scholar 

  • C.M. Palmeira A.J. Moreno V.M. Madeira (1994a) ArticleTitleInteractions of herbicides 2,4-D and dinoseb with liver mitochondrial bioenergetics Toxicol. Appl. Pharmacol. 127 50–57 Occurrence Handle1:CAS:528:DyaK2cXltVWmtLo%3D Occurrence Handle10.1006/taap.1994.1138

    Article  CAS  Google Scholar 

  • C.M. Palmeira A.J. Moreno V.M. Madeira (1994b) ArticleTitleMetabolic alterations in hepatocytes promoted by the herbicides paraquatdinoseb and 2,4-D Arch. Toxicol. 68 24–31 Occurrence Handle1:CAS:528:DyaK2cXitFSgurY%3D Occurrence Handle10.1007/s002040050027

    Article  CAS  Google Scholar 

  • D.B. Papkovsky T. O’Riordan A. Soini (2000) ArticleTitlePhosphorescent porphyrin probes in biosensors and sensitive bioassays Biochem. Soc. Trans. 28 74–77 Occurrence Handle10816102 Occurrence Handle1:CAS:528:DC%2BD3cXhsVKqtrk%3D

    PubMed  CAS  Google Scholar 

  • A. Ramanathan C. Wang S. Schreiber (2005) ArticleTitlePerturbational profiling of a cell-line model of tumorigenesis by using metabolic measurements Proc. Natl. Acad. Sci. USA 102 5992–5997 Occurrence Handle15840712 Occurrence Handle1:CAS:528:DC%2BD2MXkt1Cmu74%3D Occurrence Handle10.1073/pnas.0502267102

    Article  PubMed  CAS  Google Scholar 

  • P.O. Seglen (1976) ArticleTitlePreparation of isolated rat liver cells Methods. Cell Biol. 13 29–83 Occurrence Handle177845 Occurrence Handle1:CAS:528:DyaE28XkslCgtbY%3D Occurrence Handle10.1016/S0091-679X(08)61797-5

    Article  PubMed  CAS  Google Scholar 

  • W. Steurer S. Stadlmann K. Roberts M. Fischer R. Margreiter E. Gnaiger (1999) ArticleTitleQuality assessment of isolated pancreatic rat islets by high-resolution respirometry Transplant Proc. 31 650 Occurrence Handle10083280 Occurrence Handle1:STN:280:DyaK1M7ot1ynuw%3D%3D Occurrence Handle10.1016/S0041-1345(98)01600-5

    Article  PubMed  CAS  Google Scholar 

  • P.S. Stewart (2003) ArticleTitleDiffusion in biofilms J. Bacteriol. 185 1485–1491 Occurrence Handle12591863 Occurrence Handle1:CAS:528:DC%2BD3sXhs1Sisrs%3D Occurrence Handle10.1128/JB.185.5.1485-1491.2003

    Article  PubMed  CAS  Google Scholar 

  • D.T. Stitt M.S. Nagar T.A. Haq M.R. Timmins (2002) ArticleTitleDetermination of growth rate of microorganisms in broth from oxygen-sensitive fluorescence plate reader measurements Biotechniques 32 684 Occurrence Handle11911671 Occurrence Handle1:CAS:528:DC%2BD38XitFCntrc%3D

    PubMed  CAS  Google Scholar 

  • I.R. Sweet G. Khalil A.R. Wallen M. Steedman K.A. Schenkman J.A. Reems S.E. Kahn J.B. Callis (2002) ArticleTitleContinuous measurement of oxygen consumption by pancreatic islets Diabetes Technol. Therapuetics 4 661–671 Occurrence Handle10.1089/152091502320798303

    Article  Google Scholar 

  • W. Wang L. Upshaw D.M. Strong R.P. Roberstson J. Reems (2005) ArticleTitleIncreased oxygen consumption rates in response to high glucose detected by a novel oxygen biosensor in no-human and human islets J. Endocrinol. 185 445–455 Occurrence Handle15930171 Occurrence Handle1:CAS:528:DC%2BD2MXlsFeqtrc%3D Occurrence Handle10.1677/joe.1.06092

    Article  PubMed  CAS  Google Scholar 

  • L. Wilson-Fritch S. Nicoloro M. Chouinard M.A. Lazar P.C. Chui J. Leszyk J. Straubhaar M.P. Czech S. Corvera (2004) ArticleTitleMitochondrial Remodeling in adipose tissue associated with obesity and treatment with rosiglitazone J. Clin. Invest. 114 1281–1289 Occurrence Handle15520860 Occurrence Handle1:CAS:528:DC%2BD2cXpsFCgtr0%3D Occurrence Handle10.1172/JCI200421752

    Article  PubMed  CAS  Google Scholar 

  • M. Wodnicka R.D. Guarino J.J. Hemperly M.R. Timmins D. Stitt J.B. Pitner (2000) ArticleTitleNovel fluorescent technology platform for high throughput cytotoxicity and proliferation assays J. Biomol. Screen. 5 141–152 Occurrence Handle10894757 Occurrence Handle1:STN:280:DC%2BD3czos1arug%3D%3D Occurrence Handle10.1177/108705710000500306

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Research & Development, BD Biosciences, 2 Oak Park Dr., Bedford, MA, USA

    Laura E. Dike, Haiyan Xia & Mark R. Timmins

  2. BD Technologies, Research Triangle Park, NC, USA

    Richard D. Guarino & Sharon C. Presnell

Authors
  1. Laura E. Dike
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Haiyan Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard D. Guarino
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sharon C. Presnell
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mark R. Timmins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mark R. Timmins.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dike, L.E., Xia, H., Guarino, R.D. et al. Rapid Method for Assessing Oxygen Consumption Rate of Cells from Transient-state Measurements of Pericellular Dissolved Oxygen Concentration. Cytotechnology 49, 133–141 (2005). https://doi.org/10.1007/s10616-006-6353-3

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-006-6353-3

Keywords

Search

Navigation