ABSTRACT
To optimize program of bovine somatic nuclear transfer, we used two different enucleation procedures (by Spindle-view system & Hoechst 33342 staining), two different procedures to introduce donor nuclei (by ooplasm microinjection & electrofusion), and three different group electrofusion parameters (group 1: 1.9 kV/cm, 10 micros, two; group 2: 1.5 kV/cm, 25 micros, two; group 3: 0.6 kV/cm, 100 micros, one) to reconstruct bovine cloned embryos. The cleavation rates and blastocyst development rates of cloned embryos were used to assess the efficiency of different operational procedure. Finally, the best combination of operational procedure, that the spindle-viewer system was used for oocytes enucleating, and donor cell was electrofused into ooplasm by electrical pulse (1.9 kV/cm, 10 micros, two) to reconstruct bovine cloned embryos. Then the excellent blastocysts were transferred to fosters for producing cloned cattle 80 high-quality cloned blastocysts were transferred into 33 fosters, two cloned calves were produced. According to the results, the optimized program could be used to produce cloned cattle.
Subject(s)
Animals , Cattle , Female , Cell Nucleus , Physiology , Cloning, Organism , Embryo Transfer , Methods , Embryo, Mammalian , Cell Biology , Physiology , Microinjections , Nuclear Transfer Techniques , Oocytes , Cell Biology , PhysiologyABSTRACT
Objective To study the effects of complex polysaccharides immune milk (CPIM) on cellular and humoral immune functions in mice. Method Sixty ICR mice were divided into five experimental groups and a control group. The mice in experimental groups were given i.p. Ganoderma lucidum polysaccharide (GLP) immune milk,Lycium barbarum polysaccharides (LBP) immune milk and different doses (low,medium and high) CPIM,and the control group was given equivalently skimmed and sterilized goat's milk respectively for 15 d. The cellular and humoral immune functions and the feces were examined. Results Transformation ability of spleen lymphocytes:There were highly marked differences between all experimental groups and control group,between medium dose group of CPIM and GLB group,LBP group (P