Background
Nuclear transfer (NT) technologies offer a means for producing the genetically modified pigs necessary to develop swine models for mechanistic studies of disease processes as well as to serve as organ donors for xenotransplantation. Most previous studies have used commercial pigs as surrogates.
Method and Results
In this study, we established a cloning technique for miniature pigs by somatic cell nuclear transfer (SCNT) using Nippon Institute for Biological Science (NIBS) miniature pigs as surrogates. Moreover, utilizing this technique, we have successfully produced an α-1, 3-galactosyltransferase knockout (GalT-KO) miniature swine. Fibroblasts procured from a NIBS miniature pig fetus were injected into 1312 enucleated oocytes. The cloned embryos were transferred to 11 surrogates of which five successfully delivered 13 cloned offspring; the production efficiency was 1.0% (13/1312). In a second experiment, lung fibroblasts obtained from neonatal GalT-KO MGH miniature swine were used as donor cells and 1953 cloned embryos were transferred to 12 surrogates. Six cloned offspring were born from five surrogates, a production efficiency of 0.3% (6/1953).
Conclusions
These results demonstrate successful establishment of a miniature pig cloning technique by SCNT using NIBS miniature pigs as surrogates. To our knowledge, this is the first demonstration of successful production of GalT-KO miniature swine using miniature swine surrogates. This technique could help to ensure a stable supply of the cloned pigs through the use of miniature pig surrogates and could expand production in countries with limited space or in facilities with special regulations such as specific pathogen-free or good laboratory practice.
Most cases of ischemic heart disease and stroke occur as a result of atherosclerosis. The
purpose of this study was to produce a new Nippon Institute for Biological Science (NIBS)
miniature pig model by somatic cell nuclear transfer (SCNT) for studying atherosclerosis.
The human apolipoprotein(a) (apo(a)) genes were transfected into kidney epithelial cells
derived from a male and a female piglet. Male cells were used as donors initially, and 275
embryos were transferred to surrogates. Three offspring were delivered, and the production
efficiency was 1.1% (3/275). Serial female cells were injected into 937 enucleated
oocytes. Eight offspring were delivered (production efficiency: 0.9%) from surrogates. One
male and 2 female transgenic miniature pigs matured well. Lipoprotein(a) was found in the
male and one of the female transgenic animals. These results demonstrate successful
production of human apo(a) transgenic NIBS miniature pigs by SCNT. Our goal is to
establish a human apo(a) transgenic NIBS miniature pig colony for studying
atherosclerosis.
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