Effect of serum starvation on the efficiency of nuclear transfer using odd-eyed white cat fibroblasts.
In the present study, we compared in vitro and in vivo development of nuclear transfer (NT) embryos derived from serum-starved or non-serum-starved odd-eyed cat skin fibroblast cells. Flow cytometry analyses revealed that a higher percentage of cells were in the G0/G1 phase after serum starvation (89.3%) as compared with non-serum-starved cells (73.8%, P<0.05). Frequency of cleavage and development to the blastocyst stage was not different between the serum-starved or non-serum-starved treatment group, 67.9 and 12.5% versus 73.0 and 10.2%, respectively (P>0.05). After transfer of two to four-cell NT embryos derived from starved and non-starved fibroblasts, three of nine (33%) and one of nine (11%) recipients delivered three live male (plus, one dead) and two live male kittens, respectively. Of the five live-cloned kittens, one died from diarrhea at 3 weeks of age and the other four kittens are growing at a normal rate. The cloned kittens are blue-eyed and have functional auditory systems, including clones of the odd-eyed deaf Turkish Angora cat. Subsequent DNA analysis of nine-cat specific microsatellite loci confirmed that all of the cloned kittens were identical to the odd-eyed donor male, but a point mutation occurred in the dead fetus at the FCA 290 marker.
Yin
Xi-Jun
XJ
Department of Animal Science & Technology, College of Agriculture & Life Science, Sunchon National University, 315 Maegok-dong, Sunchon, 540-742 JeonNam Province, South Korea.
Lee
Hyo-Sang
HS
Kim
Lyoung-Ho
LH
Shin
Hyoung-Doo
HD
Kim
Nam-Hyung
NH
Kong
Il-Keun
IK
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D001835:Body Weight; D002415:Cats / Q000502:physiology*; D002453:Cell Cycle; D002460:Cell Line; D002999:Clone Cells; D003470:Culture Media; D016895:Culture Media, Serum-Free; D047108:Embryonic Development / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology / Q000502:physiology
D003470:Culture Media; D016895:Culture Media, Serum-Free