KMID : 0900920030270030233
|
|
Korean journal of Animal Reproduction 2003 Volume.27 No. 3 p.233 ~ p.240
|
|
Study on the Improvement of Bovine Somatic Cell Nuclear Transfer Technique
|
|
Lee Sang-Young Park Choon-Keun Yang Boo-Keun Kim Choung-Ik Cheong Hee-Tae
|
|
Abstract
|
|
|
This study was conducted to examine the effects of oocyte maturation period, phytohemagglutinin-P (PHA-P) treatment and activation agent on the enucleation, fusion, activation or in vitro development of bovine nuclear transfer embryos. Bovine oocytes were enucleated at 16¡24 h of in vitro maturation (IVM). Adult ear skin cells treated or non-treated with PHA-P were transferred into enucleated oocytes. Reconstituted oocytes treated or non-treated with PHA-P were fused by a pulse of 1.5 kV/cm for 30 musec. Fused oocytes were activated with a combination of calcium ionophore (A23187) and cycloheximide (CHXM) or dimethylaminopurine (DMAP), and cultured in vitro for 7¡9 days. Enucleation rate was significantly increased when oocytes were matured for 16¡18 h (70.2¡92.3%, P<0.05) compared to that of oocytes were matured for 20¡24 h (44.3¡53.4%). The location of metaphase-II plate was far off from the 1st polar body as maturation time was increased. PHA-P treatment of donor cells or reconstituted oocytes significantly improved fusion rate (P<0.05). Cleavage and blastocyst formation rates were significantly increased after activation with a combination of A23187 and DMAP (78.6% and 32.9%, respectively) compared to those of embryos activated with a combination of A23l87 and CHXM (48.5 and 15.2%, respectively). From the present result, it is suggested that high enucleation efficiency can obtained by using oocytes matured for 18 h. It also shows that PHA-P treatment can improve the fusion rate, and activation with a combination of A23187 and DMAP can enhance the embryo development.
|
|
KEYWORD
|
|
Nuclear transfer, Efficiency, In vitro development, Bovine somatic cells
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|