Study On The Establishment Of Mouse Embryonic Stem Cell Lines

PurposeTo explore the method to establish mouse embryonic stem cell (ES) line, on the basis of which, the method will offer the technique and theory support for the establishment and future study of human being ES line.MethodsTwo kinds of mouse (ICR&129) were selected in present study. Nature-fertilized blastocysts were obtained, subsequently, inner cell masses (ICMs) were separated and grafted on the layer of mouse embryonic fibroblast cells to conduct culture and passage to establish the cell lines, and then these cell lines would be identified as ES line by use of morphological observation, AKP examination, in-vitro and in-vivo differentiation test.Results1. Result on construction of Nature-fertilized ES line of mouse from 129 genetic background.:Totally 58 blastocysts were obtained in the study. After 3 to 5 days of coculture with feeder cells, inner cell masses(ICM) in 47 blastocysts were found increasing significantly. These ICMs were then separated , conducted passage and 36 primary ES colonies were obtained among them. Subsequently, separation and passage were repeated few times and finally one ES cell line was obtained by identification. At present, the ES line has developed to its 36th generation.2. Result on construction of nature-fertilized ES line fro ICR mouse: Totally 92 blastocysts were collected and 69 blastocysts of ICMs were observed increasing significantly after culture in medium. After separation and passage, there were 34 primary ES colonies present in them. Finally, no ES cell lines were obtained in the...