Postneonatal Mortality and Liver Changes in Cloned Pigs Associated with Human Tumor Necrosis Factor Receptor I-Fc and Human Heme Oxygenase-1 Overexpression

Figure 1

Demonstration of transgenesis in cloned pigs produced by SCNT. (a) Picture of cloned pigs after birth. Birth weight piglet #1: 1030 g, piglet #2: 750 g, piglet #3: 764 g, piglet #4: 640 g, piglet #5: 453 g, piglet #6: 406 g, and piglet #7: 400 g. (b) PCR analysis on genomic DNA isolated from the produced transgenic animals (#1~#7) and nontransgenic control, wild type. PCR with shTNFRI-Fc (upper panel) and HO-1 (lower panel) specific primers demonstrated the presence of the shTNFRI-Fc and hHO-1 transgene. P, plasmid control; N, negative control. (c) Resistance of fibroblasts from transgenic pigs against oxidative stress. The fibroblasts of 1 × 10⁴ cells per well from transgenic pig were stimulated with hTNF-α (20 ng/ml) and CHX (10 micrograms/ml) stimulation for 15 h and the viability was determined by CCK-8. values versus wild type pig fibroblasts (). The data shown were obtained in six replicates. (d) Cell viability of fibroblasts against apoptotic stimulation. Fibroblasts of 1 × 10⁴ cells per well from each TG pig were treated with H₂O₂ (400 μM) for 1 h. values versus wild type pig fibroblasts (). The data shown were obtained in six replicates. Columns carrying different letters are statistically different at < 0.05.

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