Cytotechnology

, Volume 2, Issue 2, pp 85–94 | Cite as

Distinct volume distribution of viable and non-viable hybridoma cells: A flow cytometric study

  • Sankar Sen
  • Friedrich Srienc
  • Wei-Shou Hu
Review

Abstract

Light scattering properties of hybridoma cells were examined with flow cytometry. Viable and dead cells form two distinct populations. The distribution of the two populations changes during a batch culture. the concentration of dead cells measured by flow cytometry correlates well to that measured by hemacytometer. The distribution based on small-angle light scattering is similar to the distribution based on volume as measured by Elzone particle counter. It thus appears that viable cells form the population with a larger mean cell volume. The results also indicate that the volume of viable cells decreases during the cultivation while that of dead cells remains relatively constant.

Key words

hybridoma cell volume cell culture flow cytometry 

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References

  1. 1.
    BrunstigA and MullaneyPF (1974) Differential light scattering from spherical mammalian cells. Biophys. J. 14: 439–453.Google Scholar
  2. 2.
    Frame KK and HU W-S: Cell volume measurement as an estimation of mammalian cell biomass. Biotechnol. Bioeng. (accepted for publication).Google Scholar
  3. 3.
    KrishanA (1975) Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J. Cell. Biol. 66: 188–193.Google Scholar
  4. 4.
    LearyJF, ToddP, WoodsUCS and JettJH (1979) Laser flow cytometric light scatter and fluorescence pulse rise-time sizing of mammalian cells. J. Histochem. Cytochem. 27: 315–320.Google Scholar
  5. 5.
    LokenMR, ParksDR and HerzenbergLA (1977) Identification of cell asymmetry and orientation by light scattering. J. Histochem. Cytochem. 25: 790–795.Google Scholar
  6. 6.
    LokenMR, StourRD and HerzenbergLA (1979) Lymphoid cell analysis and sorting. In Flow Cytometry and Sorting. MelamedMR, MullaneyPF, MendelsohnML Eds., John Wiley and Sons, NY, p. 505–528.Google Scholar
  7. 7.
    MuirheadKA, HoranPK and PosteG (1985) Flow cytometry: present and future. Biotechnology, 3: 337–355.Google Scholar
  8. 8.
    MullaneyPF, VanDillaMA, CoulterJR and DeanPN (1969) Cell sizing: a light scatter photometer for rapid volume determination. Rev. Sci. Instrum. 40: 1029.Google Scholar
  9. 9.
    SasakiDT, DumasSE and EnglemanEG (1987) Discrimination of viable and non-viable cells using propidium iodide in two color immunofluorescence. Cytometry 8: 413–420.Google Scholar
  10. 10.
    SteinkampJA (1984) Flow cytometry. Rev. Sci. Instrum. 55: 1375–1400.Google Scholar
  11. 11.
    TraganosF (1984) Flow cytometry: principles and applications. 1. Cancer Invest. 2: 149–163.Google Scholar

Copyright information

© Kluwer Academic Publishers 1989

Authors and Affiliations

  • Sankar Sen
    • 1
  • Friedrich Srienc
    • 1
    • 2
  • Wei-Shou Hu
    • 1
  1. 1.Department of Chemical Engineering and Materials ScienceUniversity of MinnesotaMinneapolisUSA
  2. 2.Institute for Advanced Studies in Biological Process TechnologyUniversity of MinnesotaSt. PaulUSA

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