Submitted January 25, 2007
Returned for revision February 16, 2007
Accepted April 27, 2007
Reproductive Technology
Buffalos (Bubalus bubalis) Cloned by Nuclear
Transfer of Somatic Cells
Deshun Shi *,
Fenghua Lu ,
Yingming Wei ,
Kuiqing Cui ,
Sufang Yang ,
Jingwei Wei ,
and
Qingyou Liu
* To whom correspondence should be addressed. E-mail: ardsshi{at}163.com.
Abstract
Cloning of buffalos (Bubalus bubalis) through nuclear transfer is a potential alternative approach in genetic improvement of buffalos. However, cloned offspring of buffalos derived from either embryonic, fetal or somatic cells has not been reported up to now. Thus, factors affecting the nuclear transfer of buffalo somatic cells were examined and the possibility of cloning buffalos was explored in this study. Treatment of buffalo fibroblasts and granulosa cells with aphidicolin plus serum starvation resulted in more cells arrested at G0/G1 phase, less proportion of cells with DNA fragmentation, and more embryos derived from these cells developed to blastocysts. In addition, a difference was also found in the development of embryos reconstructed with fetal fibroblasts from different individuals (P < 0.001). Fourty two blastocysts derived from granulosa cells and fetal fibroblasts were transferred into 21 recipient swamp buffalos, and four recipients were confirmed to be pregnant by rectal palpation on day 60 of gestation. One recipient received two embryos from fetal fibroblasts aborted on day 300 of gestation and delivered two female premature calves. Three recipients maintained pregnancy to term and delivered 3 female cloned calves after day 338 to 349 of gestation. These results indicate that buffalo embryos derived from either fetal fibroblasts or granulosa cells can develop to the term of gestation and result in newborn calves.
Key words:
Embryo •
Pregnancy •
Developmental biology •
Early development
