The invention discloses a method for improving in-vitro development efficiency of porcine cloned embryos. The method comprises the following steps of: performing
molecular identification on
bone marrow-derived mesenchymal stem cells, and establishing a method capable of identifying the purity of the cells by using a few cells through a 6-channel flow type technology; performing
gene modification on the
porcine bone marrow mesenchymal stem cells to obtain
porcine bone marrow mesenchymal stem cells for expressing multiple
reprogramming factors; and separating out positive
porcine bone marrow mesenchymal stem cells by using a fluorescent flow type
separation method, continuously culturing the positive porcine
bone marrow mesenchymal stem cells for 3 to 7 days, performing
nuclear transfer on the cultured cells serving as nuclear donor cells, and studying the influence of 4RFs-3days, 6RFs-3days, 4RFs-7days and 6RFs-7days pMSC serving as nuclear donor cells on the improvement of the development efficiency of the porcine cloned embryos. The results show that the 4RFs-7days pMSC can well promote
fission of the cloned embryos and formation of blastospheres, the cloned embryos can form a homogeneous state with uniform
cell quantity, and a foundation is laid for efficiently
cloning adult high-quality pig varieties in large scale.