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Technology for cell cycle research with unstressed steady-state cultures
Authors:Valerie S. LeBleu  Maureen Thornton  Steven R. Gonda  Charles E. Helmstetter
Affiliation:(1) Department of Biological Sciences, Florida Institute of Technology, Melbourne, FL 32901, USA;(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;(3) Midwest Research Institute-Florida Division, Palm Bay, FL 32909, USA;(4) Johnson Space Center, Houston, TX, USA
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.
Contact InformationValerie S. LeBleuEmail: Phone: +617-667-0427Fax: +617-667-2562
Keywords:Steady-state culture  Cell cycle  Continuous culture  Rotating culture
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