원문정보
초록
영어
The technique of SCNT is now well established but still remains inefficient. The in vitro development of SCNT embryos is dependent upon numerous factors including the recipient cytoplast and karyoplast. Above all, the metaphase of the second meiotic division (MII) oocytes have typically become the recipient of choice. Generally high level of MPF present in MII oocytes induces the transferred nucleus to enter mitotic division precociously and causes NEBD and PCC, which may be the critical role for nuclear reprogramming. In the present study we investigated the in vitro development and pregnancy of White-Hanwoo SCNT embryos treated with caffeine (a protein kinase phosphatase inhibitor). As results, the treatment of 10 mM caffeine for 6 h significantly increased MPF activity in bovine oocytes but does not affect the developmental competence to the blastocyst stage in bovine SCNT embryos. However, a significant increase in the mean cell number of blastocysts and the frequency of pregnant on 150 days of White-Hanwoo SCNT embryos produced using caffeine treated cytoplasts was observed. These results indicated that the recipient cytoplast treated with caffeine for a short period prior to reconstruction of SCNT embryos is able to increase the frequency of pregnancy in cow.
목차
MATERIALS & METHODS
In Vitro Oocyte Maturation
Caffeine Treatment and Parthenogenetic Activation of In Vitro Mature MII Oocytes
Oocyte Enucleation
Donor Cell Culture
Cell Fusion and Activation
Culture of Reconstructed Embryos
MPF Activity Assay
Statistical Analysis
RESULTS
MPF Activity at Different Stages of Oocyte Maturation
Determination of the Optimal Concentration of Caffein
Determination of the Optimal Period of Exposure to Caffeine
Development of NT Embryos Reconstructed using CaffeineTreated Oocytes as Cytoplast Recipients
DISCUSSION
REFERENCES
