Somatic cell cloning without micromanipulators.
Until now, micromanipulators have been regarded as indispensable for somatic cell nuclear transfer. This paper describes an improved zona-free nuclear transfer procedure with manual bisection of oocytes, selection of cytoplasts by Hoechst staining, and two-step fusion of somatic cells from primary granulosa cell cultures with two cytoplasts. Blastocyst rates in the three systems tested for zona-free embryo culture were 0%, 18%, and 36% for microdrops, well of the wells (WOW system), and microcapillaries (GO system), respectively. This simple, rapid, and inexpensive procedure may become a useful alternative to the existing techniques for somatic cell nuclear transfer for large-scale application of the technology.
Vajta
G
G
Centre for Early Human Development, Monash Institute of Reproduction and Development, Monash University, Clayton, Victoria, Australia.
Lewis
I M
IM
Hyttel
P
P
Thouas
G A
GA
Trounson
A O
AO
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism*; D019976:Cloning, Organism / Q000379:methods*; D003470:Culture Media / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology*; D013997:Time Factors
D003470:Culture Media