Abstract
The incidence of metabolic syndrome is increasing globally, as the prevalence of obesity continues to rise. However, the basic mechanisms of metabolic syndrome are not completely known yet. Therefore, animal disease models are required for the study of metabolic syndrome. The overexpression of 11 β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) in mice leads to metabolic syndrome; thus, we attempted to produce pigs with overexpression of 11β-HSD1 gene by somatic cell nuclear transfer (SCNT). However, low transgenic (TG) efficiency has been an obstacle to the production of TG pigs. A SCNT method in which somatic cells derived from TG pig are used as the nuclear donor (re-cloning method) is an effective technique for TG pig production. In this study, we attempted to increase TG efficiency by the re-cloning method. Pregnancy efficiency, production efficiency, and TG efficiency were compared with sources of donor cells (transfected TG fetal fibroblast v. TG fibroblast derived from newborn TG cloned pig). A total of 1382 and 881 TG SCNT embryos were produced from fetal fibroblast v. cloned fibroblast, and then transferred to 13 and 10 recipients. The pregnancy rate was not significantly different (30.8% v. 20.0%). Seventeen live piglets and 5 stillborn piglets were born from 4 recipients in the fetal fibroblast group, and 8 live piglets, 2 stillborn piglets, and 3 mummies were born from 2 recipients in the cloned fibroblast group. There were no significant differences in the production efficiency (3.7% v. 5.0%). All of the 13 re-cloned piglets showed reporter and target gene integration. But, of 22 fetal fibroblast-cloned piglets, reporter gene integration was confirmed in 9, but only 3 clone piglets showed reporter gene integration. Efficiency of TG was significantly increased in re-cloning group (13.6% v. 100.0%). In this study, TG efficiency of 11β-HSD1 overexpressed pigs was improved by re-cloning method. These results indicate that re-cloning is an efficient method for production of TG cloned pigs.This work was supported by a grant from the Next-Generation Bio Green 21 Program (No. PJ009563032014), Rural Development Administration, Republic of Korea.
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