Coordination between donor cell type and cell cycle stage improves nuclear cloning efficiency in cattle.
Several studies have shown that both quiescent and proliferating somatic donor cells can be fully reprogrammed after nuclear transfer (NT) and result in viable offspring. So far, however, no comparative study has conclusively demonstrated the relative importance of donor cell cycle stage on nuclear cloning efficiency. Here, we compare two different types of bovine fetal fibroblasts (BFFs) that were synchronized in G(0), G(1), and different phases within G(1). We show that for non-transgenic (non-TG) fibroblasts, serum starvation into G(0) results in a significantly higher percentage of viable calves at term than synchronization in early G(1) or late G(1). For transgenic fibroblasts, however, cells selected in G(1) show significantly higher development to calves at term and higher post-natal survival to weaning than cells in G(0). This suggests that it may be necessary to coordinate donor cell type and cell cycle stage to maximize overall cloning efficiency.
Wells
D N
DN
Reproductive Technologies, AgResearch Ltd, Ruakura Research Centre, East Street, Private Bag 3123, Hamilton, New Zealand. david.wells@agresearch.co.nz
Laible
G
G
Tucker
F C
FC
Miller
A L
AL
Oliver
J E
JE
Xiang
T
T
Forsyth
J T
JT
Berg
M C
MC
Cockrem
K
K
L'Huillier
P J
PJ
Tervit
H R
HR
Oback
B
B
D000818:Animals; D002417:Cattle / Q000196:embryology / Q000235:genetics*; D002453:Cell Cycle; D019976:Cloning, Organism; D004624:Embryo Transfer / Q000662:veterinary; D005314:Embryonic and Fetal Development; D005260:Female; D005347:Fibroblasts / Q000648:ultrastructure; D016193:G1 Phase; D016195:G2 Phase; D008938:Mitosis; D053652:Nuclear Transfer Techniques; D011247:Pregnancy; D016192:Resting Phase, Cell Cycle