Flow cytometric cell-cycle analysis of cultured fibroblasts from the giant panda, Ailuropoda melanoleuca L.

被引:20
作者
Han, ZM
Chen, DY
Li, JS
Sun, QY
Wang, PY
Du, J
Zhang, HM
机构
[1] Chinese Acad Sci, Inst Zool, State Key Lab Reprod Biol, Beijing 100080, Peoples R China
[2] Beijing Normal Univ, Coll Life Sci, Beijing 100875, Peoples R China
[3] China Res & COnservat Ctr Giant Panda, Wolong Nat Reserve, Karaikkudi 623006, Tamil Nadu, India
关键词
fibroblast; giant panda; cell cycle; flow cytometry; preparation for cloning; NUCLEAR TRANSFER; CAPRA-HIRCUS; CLONED PIGS; ADULT; FETAL; CLONING; GENERATION; CATTLE; GOATS; MICE;
D O I
10.1016/S1065-6995(02)00353-0
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
In animal cloning, it is generally believed that the inactive diploid G(0) or G(1) stage of the cell cycle is beneficial to initiate cell-cycle coordination and reprogramming following transfer of the donor nucleus. Previous experiments have demonstrated that serum starvation results in quiescent cell stage. Some experiments show that the majority of cells in a fully confluent cell culture are also in an inactive G(1) stage. In order to provide more G(0)/G(1) stage cells for giant panda cloning, we carried out a flow cytometric analysis of the cell cycle of fibroblasts from the abdominal muscle of a giant panda at different passage numbers under different growth conditions, and after different periods of serum starvation. The percentage of G(0) + G(1), stage cells differed significantly under different growth conditions. Serum starvation effectively increased the percentage of G(0) + G(1) stage cells, and the cell cycle characteristics following serum starvation for varying periods of time differed with this and the initial confluency of the cultures. The data should help in choosing the optimal stage for preparing donor cells as well as increasing the potential cloning efficiency in our study of giant panda cloning. (C) 2003 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:349 / 353
页数:5
相关论文
共 19 条
[1]   Production of goats by somatic cell nuclear transfer [J].
Baguisi, A ;
Behboodi, E ;
Melican, DT ;
Pollock, JS ;
Destrempes, MM ;
Cammuso, C ;
Williams, JL ;
Nims, SD ;
Porter, CA ;
Midura, P ;
Palacios, MJ ;
Ayres, SL ;
Denniston, RS ;
Hayes, ML ;
Ziomek, CA ;
Meade, HM ;
Godke, RA ;
Gavin, WG ;
Overström, EW ;
Echelard, Y .
NATURE BIOTECHNOLOGY, 1999, 17 (05) :456-461
[2]   Production of cloned pigs from in vitro systems [J].
Betthauser, J ;
Forsberg, E ;
Augenstein, M ;
Childs, L ;
Eilertsen, K ;
Enos, J ;
Forsythe, T ;
Golueke, P ;
Jurgella, G ;
Koppang, R ;
Lesmeister, T ;
Mallon, K ;
Mell, G ;
Misica, P ;
Pace, M ;
Pfister-Genskow, M ;
Strelchenko, N ;
Voelker, G ;
Watt, S ;
Thompson, S ;
Bishop, M .
NATURE BIOTECHNOLOGY, 2000, 18 (10) :1055-1059
[3]   The giant panda (Ailurapoda melanoleuca) somatic nucleus can dedifferentiate in rabbit ooplasm and support early development of the reconstructed egg [J].
Chen, DY ;
Sun, QY ;
Liu, JL ;
Li, GP ;
Lian, L ;
Wang, MK ;
Han, ZM ;
Song, XF ;
Li, JS ;
Sun, Q ;
Chen, YC ;
Zhang, YP ;
Ding, B .
SCIENCE IN CHINA SERIES C-LIFE SCIENCES, 1999, 42 (04) :346-353
[4]  
Han ZM, 2001, IN VITRO CELL DEV-AN, V37, P644
[5]   Development rates of male bovine nuclear transfer embryos derived from adult and fetal cells [J].
Hill, JR ;
Winger, QA ;
Long, CR ;
Looney, CR ;
Thompson, JA ;
Westhusin, ME .
BIOLOGY OF REPRODUCTION, 2000, 62 (05) :1135-1140
[6]   Eight calves cloned from somatic cells of a single adult [J].
Kato, Y ;
Tani, T ;
Sotomaru, Y ;
Kurokawa, K ;
Kato, JY ;
Doguchi, H ;
Yasue, H ;
Tsunoda, Y .
SCIENCE, 1998, 282 (5396) :2095-2098
[7]   Generation of dwarf coat (Capra hircus) clones following nuclear transfer with transfected and nontransfected fetal fibroblasts and in vitro-matured oocytes [J].
Keefer, CL ;
Baldassarre, H ;
Keyston, R ;
Wang, B ;
Bhatia, B ;
Bilodeau, AS ;
Zhou, JF ;
Leduc, M ;
Downey, BR ;
Lazaris, A ;
Karatzas, CN .
BIOLOGY OF REPRODUCTION, 2001, 64 (03) :849-856
[8]   Six cloned calves produced from adult fibroblast cells after long-term culture [J].
Kubota, C ;
Yamakuchi, H ;
Todoroki, J ;
Mizoshita, K ;
Tabara, N ;
Barber, M ;
Yang, XZ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (03) :990-995
[9]   Pig cloning by microinjection of fetal fibroblast nuclei [J].
Onishi, A ;
Iwamoto, M ;
Akita, T ;
Mikawa, S ;
Takeda, K ;
Awata, T ;
Hanada, H ;
Perry, ACF .
SCIENCE, 2000, 289 (5482) :1188-1190
[10]   Cloned pigs produced by nuclear transfer from adult somatic cells [J].
Polejaeva, IA ;
Chen, SH ;
Vaught, TD ;
Page, RL ;
Mullins, J ;
Ball, S ;
Dai, YF ;
Boone, J ;
Walker, S ;
Ayares, DL ;
Colman, A ;
Campbell, KHS .
NATURE, 2000, 407 (6800) :86-90