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Technology for cell cycle research with unstressed steady-state cultures

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Abstract

A culture system for performing cell cycle analyses on cells in undisturbed steady-state populations was designed and tested. In this system, newborn cells are shed continuously from an immobilized, perfused culture rotating about the horizontal axis. As a result of this arrangement, the number of newborn cells released into the effluent medium each generation is identical to the number of cells residing in the immobilized population, indicating that one of the two new daughter cells is shed at each cell division. Thus, the immobilized cells constitute a continuous, steady-state culture because the concentrations, locations and microenvironments of the cells in the culture vessel do not vary with time. In tests with mouse L1210 lymphocytic leukemia cells, about 108 newborn cells were produced per day. This new culture system enables a multiplicity of cell cycle analyses on large numbers of cells assured to be from populations in steady-state growth.

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Notes

  1. American Type Culture Collection, Manassas, VA, 20110, USA.

  2. Invitrogen, Carlsbad, CA, 92008.

  3. Sigma-Aldrich Co., St. Louis, MO, 63178, USA.

  4. Millipore Corp., Bedford, MA, 01821, USA.

  5. Geotech Inc., Denver, CO, 80207, USA.

  6. Celdyne Corp., Houston, TX, 77058, USA.

  7. Beckman Coulter Inc., Fullerton, CA, 92834, USA.

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Acknowledgements

Supported, in part, by NASA grants NAG8-1582 and NAG2-1508 to CEH.

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Authors and Affiliations

  1. Department of Biological Sciences, Florida Institute of Technology, Melbourne, FL, 32901, USA

    Valerie S. LeBleu & Charles E. Helmstetter

  2. Biological and Biomedical Sciences Program, Division of Matrix Biology, Kalluri Laboratory, Harvard Medical School, BIDMC, DANA 515, 330 Brookline Ave, Boston, MA, 02446, USA

    Valerie S. LeBleu

  3. Midwest Research Institute-Florida Division, Palm Bay, FL, 32909, USA

    Maureen Thornton

  4. Johnson Space Center, Houston, TX, USA

    Steven R. Gonda

Authors
  1. Valerie S. LeBleu
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  2. Maureen Thornton
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  3. Steven R. Gonda
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  4. Charles E. Helmstetter
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Correspondence to Valerie S. LeBleu.

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LeBleu, V.S., Thornton, M., Gonda, S.R. et al. Technology for cell cycle research with unstressed steady-state cultures. Cytotechnology 51, 149–157 (2006). https://doi.org/10.1007/s10616-006-9024-5

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  • DOI: https://doi.org/10.1007/s10616-006-9024-5

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