Skip to main content
NCBI home page
Cytotechnology logoLink to Cytotechnology
. 1999 Jul;30(1-3):121–132. doi: 10.1023/A:1008039011960

Analysis of CHO-K1 cell growth in a fixed bed bioreactor using magnetic resonance spectroscopy and imaging

Peter E Thelwall 1, Kevin M Brindle 1
PMCID: PMC3449953  PMID: 19003361

Abstract

Non-invasive magnetic resonance imaging and spectroscopy techniques have been used to monitor the growth and distribution of Chinese hamster ovary K1 cells growing in a fixed bed bioreactor composed of macroporous carriers. Diffusion-weighted 1H magnetic resonance spectroscopy was used to monitor the volume fraction of the bioreactor occupied by the cells and diffusion-weighted 1H magnetic resonance imaging was used to map cell distribution. The imaging measurements demonstrated that cell growth in the bioreactor was heterogeneous, with the highest cell densities being found at the surface of the carriers. The increase in the volume fraction occupied by the cells during cell growth showed a close correlation with bioreactor ATP content measured using 31P magnetic resonance spectroscopy. These magnetic resonance measurements, in conjunction with measurements of bioreactor glucose consumption, allowed estimation of the specific glucose consumption rate. This declined during the culture, in parallel with medium glucose concentration.

Keywords: bioreactor, cell volume, imaging, magnetic resonance

Full Text

The Full Text of this article is available as a PDF (126.8 KB).

References

  1. Banik GG, Heath CA. Hybridoma growth and antibody production as a function of cell density and specific growth rate in perfusion culture. Biotechnol Bioeng. 1995;48:289–300. doi: 10.1002/bit.260480315. [DOI] [PubMed] [Google Scholar]
  2. Barry JA, McGovern KA, Lien Y-HH, Ashmore B, Gillies RJ. Dimethyl methylphosphonate (DMMP): A 31P nuclear magnetic resonance spectroscopic probe of intracellular volume in mammalian cell cultures. Biochemistry. 1993;32:4665–4670. doi: 10.1021/bi00068a026. [DOI] [PubMed] [Google Scholar]
  3. Borys MC, Linzer DIH, Papoutsakis ET. Culture pH affects expression rates and glycosylation of recombinant mouse placental lactogen proteins by Chinese Hamster Ovary (CHO) cells. Bio/Technology. 1993;11:720–724. doi: 10.1038/nbt0693-720. [DOI] [PubMed] [Google Scholar]
  4. Blute T, Gillies RJ, Dale BE. Cell density measurements in hollow fibre bioreactors. Biotechnol Prog. 1988;4:202–209. doi: 10.1002/btpr.5420040403. [DOI] [Google Scholar]
  5. Brindle KM. Investigating the performance of intensive mammalian cell bioreactor systems using magnetic resonance imaging and spectroscopy. Biotech Genetic Eng Rev. 1998;15:499–520. doi: 10.1080/02648725.1998.10647967. [DOI] [PubMed] [Google Scholar]
  6. Callies R, Jackson ME, Brindle KM. Measurements of the growth and distribution of mammalian cells in a hollow-fiber bioreactor using nuclear magnetic resonance imaging. Bio/Technology. 1994;12:75–78. doi: 10.1038/nbt0194-75. [DOI] [PubMed] [Google Scholar]
  7. Chresand TJ, Gillies RJ, Dale BE. Optimum fiber spacing in a hollow fiber bioreactor. Biotechnol Bioeng. 1988;32:983–992. doi: 10.1002/bit.260320806. [DOI] [PubMed] [Google Scholar]
  8. Donoghue C, Brideau M, Newcomer P, Pangrle B, DiBiasio D, Walsh E, Moore S. Use of magnetic resonance imaging to analyse the performance of hollow fibre bioreactors. Ann NY Acad Sci. 1992;665:285–300. doi: 10.1111/j.1749-6632.1992.tb42592.x. [DOI] [PubMed] [Google Scholar]
  9. Drury DD, Dale BE, Gillies RJ. Oxygen transfer properties of a bioreactor for use within a nuclear magnetic resonance spectrometer. Biotechnol Bioeng. 1988;32:966–974. doi: 10.1002/bit.260320804. [DOI] [PubMed] [Google Scholar]
  10. Fernandes EJ. Nuclear magnetic resonance spectroscopy and imaging. In: Willaert RG, Baron GV, de Backer L, editors. Immobilised living cell systems. Chichester, UK: John Wiley & Sons; 1996. pp. 117–146. [Google Scholar]
  11. Hayter PM, Curling EMA, Baines AJ, Jenkins N, Salmon I, Strange PG, Tong JM, Bull AT. Glucose-limited chemostat culture of Chinese Hamster Ovary cells producing recombinant human interferon-γ. Biotechnol Bioeng. 1992;39:327–335. doi: 10.1002/bit.260390311. [DOI] [PubMed] [Google Scholar]
  12. Gillies RJ, MacKenzie NE, Dale BE. Analyses of bioreactor performance by nuclear magnetic resonance spectroscopy. Bio/Technology. 1989;7:50–54. doi: 10.1038/nbt0189-50. [DOI] [Google Scholar]
  13. Gilles RJ, Scherer PG, Raghunand N, Okerland LS, Martinez-Zaguilan R, Hersterberg L, Dale BE. Iteration of hybridoma growth and productivity in hollow fibre bioreactors using 31P NMR. Magn Reson Med. 1991;18:181–192. doi: 10.1002/mrm.1910180118. [DOI] [PubMed] [Google Scholar]
  14. Hammer BE, Heath CA, Mirer SD, Belfort G. Quantitative flow measurements in bioreactors by nuclear magnetic resonance imaging. Bio/Technology. 1990;8:327–330. doi: 10.1038/nbt0490-327. [DOI] [PubMed] [Google Scholar]
  15. Hubbell JA, Langer R. Tissue engineering. Chem Eng News. 1995;73:42–54. [Google Scholar]
  16. Jauregui HO, Chowdhury NR, Chowdhury JR. Use of mammalian liver cells for artificial liver support. Cell Transplantation. 1996;5:353–367. doi: 10.1016/0963-6897(95)02020-9. [DOI] [PubMed] [Google Scholar]
  17. Knazek RA, Gullino PM, Kholer PO, Dedrick RL. Cell growth on artificial capillaries: an approach to tissue growth in vitro. Science. 1972;178:65–67. doi: 10.1126/science.178.4056.65. [DOI] [PubMed] [Google Scholar]
  18. Knight P. Hollow fiber bioreactors for mammalian cell culture. Bio/Technology. 1989;7:459–461. doi: 10.1038/nbt0589-459. [DOI] [Google Scholar]
  19. Langer R, Vacanti JP. Tissue engineering. Science. 1993;260:920–926. doi: 10.1126/science.8493529. [DOI] [PubMed] [Google Scholar]
  20. Lin AA, Kimura R, Miller WM. Production of tPA in recombinant CHO cells under oxygen-limited conditions. Biotechnol Bioeng. 1993;42:339–350. doi: 10.1002/bit.260420311. [DOI] [PubMed] [Google Scholar]
  21. Ljunggren J, Häggström L. Catabolic control of hybridoma cells by glucose and glutamine limited fed batch cultures. Biotechnol Bioeng. 1994;44:808–818. doi: 10.1002/bit.260440706. [DOI] [PubMed] [Google Scholar]
  22. Lund P. In: Methods of Enzymatic Analysis. Bergmeyer HU, editor. New York: Academic Press; 1985. pp. 357–363. [Google Scholar]
  23. Mancuso A, Fernandes EJ, Blanch HW, Clarke DS. A nuclear magnetic resonance technique for determining hybridoma cell concentration in hollow fibre bioreactors. Bio/Technology. 1990;8:1282–1285. doi: 10.1038/nbt1290-1282. [DOI] [PubMed] [Google Scholar]
  24. Neermann J, Wagner R. Comparative analysis of glucose and glutamine metabolism in transformed mammalian cell lines, insect and primary liver cells. J Cell Phys. 1996;166:152–169. doi: 10.1002/(SICI)1097-4652(199601)166:1<152::AID-JCP18>3.0.CO;2-H. [DOI] [PubMed] [Google Scholar]
  25. Noll T, Biselli M, Wandrey C. On-line biomass monitoring of immobilised hybridoma cells by dielectrical measurements. In: Carrondo MJT, Griffiths B, Moreira JLP, editors. Animal Cell Technology. The Netherlands: Kluwer Academic Publishers; 1996. pp. 289–294. [Google Scholar]
  26. Ong CP, Pörtner R, Märkl H, Yamazsaki Y, Yasuda K, Matsumura M. High density cultivation of hybridoma in charged porous carriers. J Biotechnol. 1994;34:259–268. doi: 10.1016/0168-1656(94)90061-2. [DOI] [PubMed] [Google Scholar]
  27. Piret JM, Cooney CL. Mammalian cell and protein distributions in ultrafiltration bioreactors. Biotechnol Bioeng. 1990;36:902–910. doi: 10.1002/bit.260360905. [DOI] [PubMed] [Google Scholar]
  28. Piret JM, Cooney CL. Model of oxygen transport limitations in hollow fibre bioreactors. Biotechnol Bioeng. 1990;37:80–92. doi: 10.1002/bit.260370112. [DOI] [PubMed] [Google Scholar]
  29. Seewoster T, Lehmann J. Cell size distribution as a parameter for the predetermination of exponential growth during repeated batch cultivation of CHO cells. Biotechnol Bioeng. 1997;55:793–797. doi: 10.1002/(SICI)1097-0290(19970905)55:5<793::AID-BIT9>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  30. van Zijl PCM, Moonen CTW, Faustino P, Pekar J, Kaplan O, Cohen JS. Complete separation of intracellular and extracellular information in NMR spectra of perfused cells by diffusion-weighted spectroscopy. Proc Natl Acad Sci USA. 1991;88:3228–3232. doi: 10.1073/pnas.88.8.3228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wiesmann R, Maier ST, Marx U, Buchholz R. Characterisation of oxygen transfer in a membrane-aerated hollow-fibre bioreactor using modified microcoaxial needle electrodes. Appl Microbiol Biotechnol. 1994;41:531–536. doi: 10.1007/s002530050185. [DOI] [Google Scholar]
  32. Williams SNO, Callies RM, Brindle KM. Mapping of oxygen tension and cell distribution in a hollow-fiber bioreactor using magnetic resonance imaging. Biotechnol Bioeng. 1997;58:56–61. doi: 10.1002/(SICI)1097-0290(19971005)56:1<56::AID-BIT6>3.0.CO;2-U. [DOI] [PubMed] [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES