Because the physiological characteristics and tissue structure of pigs are very similar to humans, the research on induced pluripotent stem cells (iPS) of large animals such as pigs has attracted the attention of scientists all over the world. However, due to the current lack of understanding of the mechanism of iPS induction, the induction of pig iPS cells obtained worldwide has not been successful until the production of cloned pigs. With the efforts of Dr. Lai Liangxue from the Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Dr. Lei Xiao from Zhejiang University, and Dr. Du Yutao from the BGI Institute of Genetics, this problem has been solved, marking China's achievements in the study of iPS . The research results were published online in the international journal Cell Research on December 18, 2012.
This research was conducted under the leadership of Academician Li Ning of China Agricultural University and assembled more than 10 research units across the country. Due to the current lack of understanding of the mechanism of iPS induction, the induced pig iPS cells produced cloned pigs after many attempts (over 100 cloned embryo transfer experiments), which has not been successful before. In the research, the team found that the expression of foreign genes and epigenetics may be the main reasons affecting the development of iPS cloned embryos. The team of Lai Liangxue differentiated the porcine iPS cells obtained from Xiao Lei's laboratory for 4-6 days. On the one hand, the iPS cells were withdrawn from the rapid cell cycle. On the other hand, nuclear transfer was performed after the expression of foreign genes decreased. Later, the rate of blastocyst development in vitro increased from 5% to about 20%. By transplanting these differentiated iPS cell cloned embryos to surrogate receptors, Lai Liangxue's research team took the lead in obtaining a healthy and active cloned piglet in 2011. Subsequently, Dr. Du Yutao's group used deacetylation inhibitors for pig iPS, and then obtained 4 iPS cloned pigs in 2012 using manual cloning.
Due to the unsuccessful establishment of embryonic stem cell lines, it has been very difficult to target genes in large animals. The emergence of iPS provides new hope for gene targeting in large animals. After obtaining gene-targeted iPS cells, on the one hand, chimera technology can be used to obtain reproductive chimeric animals, and genetically modified animals can be obtained through mating; on the other hand, these gene-targeted iPS cells can be used as nuclear donors by nuclear transfer Technology to obtain genetically modified cloned animals.
This research is the first time in the world that a living iPS cloned pig has been obtained, which opens up a technical route for using iPS to achieve gene targeting in large animals.
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