title: Preliminary study on somatic cell nuclear transfer in rabbits in Thailand.
abstract: The objective of the study was to develop the somatic nuclear transfer technique by using rabbits as the model. The oocyte recipients aged 16 h post coitus were collected surgically from 20 superovulated rabbit doe with 28 and 40 mg Follicle Stimulating Hormone (FSH) after mating with a vasectomized male. The metaphase II plate and 1st polar body of oocyte was later aspirated by enucleated micropipette under an inverted microscope. A single donor cell; cumulus cell or cultured or frozen fibroblast cell from passage 1 to 9 were transferred to enucleated oocyte and fused with triple DC pulses, 3.2 kv, 20 micros. The fused embryos were cultivated in TCM 199 NaHCO3 + 10 per cent fetal calf serum (FCS) for 4 days. The cleavage rate (2-cell stage) was 37.2 per cent (32/86) from eight experiments, and 18.8 per cent (6/32) developed to the early morula stage. This study also indicated that the enucleation pipette and the somatic cell type influenced the success.
authors.affiliation: Department of Obstetrics and Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Mongkol
authors.initials: M
authors.lastname: Techakumphu
authors.forename: Pranee
authors.initials: P
authors.lastname: Numchaisrika
authors.forename: Somchai
authors.initials: S
authors.lastname: Suwajanakorn
authors.forename: Kamtorn
authors.initials: K
authors.lastname: Pruksananonda
authors.forename: Wisut
authors.initials: W
authors.lastname: Boonkasemsanti
authors.forename: Vichuda
authors.initials: V
authors.lastname: Ahnonkitpanit
authors.forename: Pramuan
authors.initials: P
authors.lastname: Virutamasen
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary; D018920:Coculture Techniques; D005314:Embryonic and Fetal Development / Q000502:physiology*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D008297:Male; D023421:Models, Animal; D009865:Oocytes / Q000254:growth & development / Q000637:transplantation*; D011817:Rabbits; D012680:Sensitivity and Specificity