Flow cytometric cell cycle analysis of cultured brown bear fibroblast cells

被引:12
作者
Caamano, J. N. [1 ]
Rodriguez, A. [1 ]
Salas, A. [2 ]
Munoz, M. [1 ]
Diez, C. [1 ]
Prather, R. S. [3 ]
Gomez, E. [1 ]
机构
[1] SERIDA, Area Genet & Reprod, Gijon 33203, Principado Astu, Spain
[2] Univ Oviedo, Serv Cientificotecn, E-33006 Oviedo, Asturias, Spain
[3] Univ Missouri, Div Anim Sci, Columbia, MO 65211 USA
关键词
Brown bear; Cell cycle; Flow cytometry; DMSO; Roscovitine;
D O I
10.1016/j.cellbi.2008.02.005
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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 mu M concentration. Interestingly, the G2/M stage significantly increased at 30 and 50 mu M compared to the control and 15 mu M (P<0.02). The cell cycle of brown bear adult fibroblast cells can be successfully synchronized under a variety of culture conditions. (C) 2008 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:855 / 859
页数:5
相关论文
共 31 条
  • [1] The cyclin-dependent kinase inhibitors olomoucine and roscovitine arrest human fibroblasts in G1 phase by specific inhibition of CDK2 kinase activity
    Alessi, F
    Quarta, S
    Savio, M
    Riva, F
    Rossi, L
    Stivala, LA
    Scovassi, AI
    Meijer, L
    Prosperi, E
    [J]. EXPERIMENTAL CELL RESEARCH, 1998, 245 (01) : 8 - 18
  • [2] Flow cytometric cell cycle analysis of cultured porcine fetal fibroblast cells
    Boquest, AC
    Day, BN
    Prather, RS
    [J]. BIOLOGY OF REPRODUCTION, 1999, 60 (04) : 1013 - 1019
  • [3] CAAMANO JN, 2005, REPROD FERT DEVELOP, V2, P245
  • [4] Campbell K H, 1996, Rev Reprod, V1, P40, DOI 10.1530/ror.0.0010040
  • [5] EFFECT OF DONOR CELL-CYCLE STAGE ON CHROMATIN AND SPINDLE MORPHOLOGY IN NUCLEAR TRANSPLANT RABBIT EMBRYOS
    COLLAS, P
    PINTOCORREIA, C
    DELEON, FAP
    ROBL, JM
    [J]. BIOLOGY OF REPRODUCTION, 1992, 46 (03) : 501 - 511
  • [6] Enhanced survivability of cloned calves derived from roscovitine-treated adult somatic cells
    Gibbons, J
    Arat, S
    Rzucidlo, J
    Miyoshi, K
    Waltenburg, R
    Respess, D
    Venable, A
    Stice, S
    [J]. BIOLOGY OF REPRODUCTION, 2002, 66 (04) : 895 - 900
  • [7] Nuclear transfer in cats and its application
    Gomez, M. C.
    Pope, C. E.
    Dresser, B. L.
    [J]. THERIOGENOLOGY, 2006, 66 (01) : 72 - 81
  • [8] Nuclear transfer of synchronized African wild cat somatic cells into enucleated domestic cat oocytes
    Gómez, MC
    Jenkins, JA
    Giraldo, A
    Harris, RF
    King, A
    Dresser, BL
    Pope, CE
    [J]. BIOLOGY OF REPRODUCTION, 2003, 69 (03) : 1032 - 1041
  • [9] Flow cytometric cell-cycle analysis of cultured fibroblasts from the giant panda, Ailuropoda melanoleuca L.
    Han, ZM
    Chen, DY
    Li, JS
    Sun, QY
    Wang, PY
    Du, J
    Zhang, HM
    [J]. CELL BIOLOGY INTERNATIONAL, 2003, 27 (04) : 349 - 353
  • [10] Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger (Panthera tigris altaica)
    Hashem, Md. Abul
    Bhandari, Dilip P.
    Kang, Sung Keun
    Lee, Byeong Chun
    [J]. MOLECULAR REPRODUCTION AND DEVELOPMENT, 2007, 74 (04) : 403 - 411