Flow cytometric cell cycle analysis of cultured brown bear fibroblast cells

Authors

  • J.N. Caamaño,

    Corresponding author
    1. Area de Genética y Reproducción, SERIDA, Camino de los Claveles 604, 33203 Gijón, Principado de Asturias, Spain
    Search for more papers by this author
  • A. Rodriguez,

    1. Area de Genética y Reproducción, SERIDA, Camino de los Claveles 604, 33203 Gijón, Principado de Asturias, Spain
    Search for more papers by this author
    • Tel.: +34 985 19 53 00; fax: +34 985 19 53 10.

  • A. Salas,

    1. Servicios Científico-Técnicos, Universidad de Oviedo, c/Julián Clavería, 33006 Oviedo, Asturias, Spain
    Search for more papers by this author
    • Tel.: +34 985 10 36 60.

  • M. Muñoz,

    1. Area de Genética y Reproducción, SERIDA, Camino de los Claveles 604, 33203 Gijón, Principado de Asturias, Spain
    Search for more papers by this author
    • Tel.: +34 985 19 53 00; fax: +34 985 19 53 10.

  • C. Diez,

    1. Area de Genética y Reproducción, SERIDA, Camino de los Claveles 604, 33203 Gijón, Principado de Asturias, Spain
    Search for more papers by this author
    • Tel.: +34 985 19 53 00; fax: +34 985 19 53 10.

  • R.S. Prather,

    1. Division of Animal Science, University of Missouri, Columbia, MO 65211, USA
    Search for more papers by this author
    • Tel.: +1 573 882 7446.

  • E. Gómez

    1. Area de Genética y Reproducción, SERIDA, Camino de los Claveles 604, 33203 Gijón, Principado de Asturias, Spain
    Search for more papers by this author
    • Tel.: +34 985 19 53 00; fax: +34 985 19 53 10.


Corresponding author. Tel.: +34 985 19 53 00; fax: +34 985 19 53 10. jncaamano@serida.org airodriguez@serida.org ana@spi.uniovi.es mmunoz@serida.org mcdiez@serida.org PratherR@missouri.edu egomez@serida.org

Abstract

The aim of this study was to assess by flow cytometry the cell cycle of brown bear fibroblast cells cultured under different growth conditions. Skin biopsies were taken in Cantabria (Spain) from a live, anaesthetized brown bear. DNA analysis was performed by flow cytometry following cell DNA staining with propidium iodide. Serum starvation increased (P < 0.01) the percentage of G0/G1 phase cells (92.7 ± 0.86) as compared to cycling cells (39.7 ± 0.86) or cells cultured to confluency (87.3 ± 0.86). DMSO included for 48 h in the culture significantly increased (P < 0.01) the percentage of G0/G1 phase of the cell cycle at all concentrations used and decreased percentages of S phase in a dose-dependent fashion. Roscovitine increased the G0/G1 phase of the cell cycle (P < 0.01) at 15 μM concentration. Interestingly, the G2/M stage significantly increased at 30 and 50 μM compared to the control and 15 μM (P < 0.02). The cell cycle of brown bear adult fibroblast cells can be successfully synchronized under a variety of culture conditions.

Ancillary