Design and application of NMR-compatible bioreactor circuits for extended perfusion of high-density mammalian cell cultures.

R. J. Gillies, Jean-Philippe Galons, K. A. McGovern, P. G. Scherer, Y. H. Lien, C. Job, R. Ratcliff, F. Chapa, S. Cerdan, B. E. Dale

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

MR spectroscopy of cultured cells allows non-invasive analyses of the metabolism of cells with specific phenotypes under defined conditions. This technique can be used to investigate the intracellular metabolism of cells or extended to critically evaluate phenomena observed by in vivo MRS. In this paper, a cell maintenance system is described which allows MR analyses with unparalleled spectral resolution, S/N and stability. This system consists of a 25 mm diameter hollow fiber bioreactor and a supporting circuit. The hollow fiber reactor was chosen because it yields a large filling factor which can be perfused through defined volumes. The fibers were 300 microns diameter microporous (0.2 micron) cellulose acetate/cellulose nitrate membranes with high porosity, which allow bulk convective flow throughout the extracapillary space. This flow (Starling flow) is necessary to disrupt steady-state gradients in substrates and waste products. In many respects, the design of the supporting circuit is more important than the bioreactor itself, since it provides the reactor with the proper chemical and physical environment. Hence, this circuit can be applied to a variety of bioreactor configurations. The circuit consists of a hollow fiber oxygenator and a bleed-and-feed system housed in a temperature-controlled cabinet. Culture of mammalian cells in this reactor yields 31P spectra which have excellent spectral and temporal resolution. At confluence, endogenous 31P line widths were typically < 10 Hz (at 162 MHz) and well resolved spectra were obtained in < 30 s.

Original languageEnglish (US)
Pages (from-to)95-104
Number of pages10
JournalNMR in Biomedicine
Volume6
Issue number1
StatePublished - Jan 1993

Fingerprint

Bioreactors
Cell culture
Cell Culture Techniques
Perfusion
Nuclear magnetic resonance
Networks (circuits)
Fibers
Metabolism
Waste Products
Oxygenators
Starlings
Collodion
Porosity
Cells
Cultured Cells
Spectral resolution
Magnetic Resonance Spectroscopy
Maintenance
Linewidth
Phenotype

ASJC Scopus subject areas

  • Biophysics
  • Spectroscopy
  • Radiology Nuclear Medicine and imaging
  • Radiological and Ultrasound Technology

Cite this

Gillies, R. J., Galons, J-P., McGovern, K. A., Scherer, P. G., Lien, Y. H., Job, C., ... Dale, B. E. (1993). Design and application of NMR-compatible bioreactor circuits for extended perfusion of high-density mammalian cell cultures. NMR in Biomedicine, 6(1), 95-104.

Design and application of NMR-compatible bioreactor circuits for extended perfusion of high-density mammalian cell cultures. / Gillies, R. J.; Galons, Jean-Philippe; McGovern, K. A.; Scherer, P. G.; Lien, Y. H.; Job, C.; Ratcliff, R.; Chapa, F.; Cerdan, S.; Dale, B. E.

In: NMR in Biomedicine, Vol. 6, No. 1, 01.1993, p. 95-104.

Research output: Contribution to journalArticle

Gillies, RJ, Galons, J-P, McGovern, KA, Scherer, PG, Lien, YH, Job, C, Ratcliff, R, Chapa, F, Cerdan, S & Dale, BE 1993, 'Design and application of NMR-compatible bioreactor circuits for extended perfusion of high-density mammalian cell cultures.', NMR in Biomedicine, vol. 6, no. 1, pp. 95-104.
Gillies, R. J. ; Galons, Jean-Philippe ; McGovern, K. A. ; Scherer, P. G. ; Lien, Y. H. ; Job, C. ; Ratcliff, R. ; Chapa, F. ; Cerdan, S. ; Dale, B. E. / Design and application of NMR-compatible bioreactor circuits for extended perfusion of high-density mammalian cell cultures. In: NMR in Biomedicine. 1993 ; Vol. 6, No. 1. pp. 95-104.
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