Preliminary study on somatic cell nuclear transfer in rabbits in Thailand
Techakumphu, M.; Numchaisrika, P.; Suwajanakorn, S.; Pruksananonda, K.; Boonkasemsanti, W.; Ahnonkitpanit, V.; Virutamasen, P.
Journal of the Medical Association of Thailand 86(3): 276-281
2003
ISSN/ISBN: 0125-2208 PMID: 12757069 Document Number: 9657
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.
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