title: Production of Cloned Wuzhishan Miniature Pigs and Application for Alloxan Toxicity Test.
abstract: Wuzhishan miniature pig is one of the four most important pig breeds in China and has many major economic characteristics. Herein, we successfully used SCNT to clone Wuzhishan miniature pig. First, ear fibroblasts were isolated from a 2-year-old female Wuzhishan miniature piglet to be used as the donor cell. Second, good-quality COCs were selected from ovaries obtained from pigs at a local slaughterhouse and cultured. Mature eggs with the first polar body and ear fibroblasts were applied SCNT. Lastly, we in total produced 12 piglets with 7 piglets surviving to adults. Next, we used these pigs to test alloxan toxicity and to build T I D diabetes type. We know that diabetes mellitus is a chronic heterogeneous metabolic disease characterized by a high blood glucose level and abnormal insulin secretion. In this study, T I D (type I diabetes) was experimentally induced in cloned Wuzhishan miniature pigs with alloxan. In brief, an intravenous injection of alloxan (group B: 170 mg/kg, n = 3) was administered to pigs weighing between 27 and 39 kg. Sterile saline was administered to control pigs (n = 3). We determined the glycometabolism related index, performed an intravenous glucose tolerance test, and carried out immunohistochemistry experiments. There were no significant differences in body weight, blood glucose, and serum insulin in all groups, before treatment. The level of blood glucose was significantly higher (P < 0.05) in group B (12.18 ± 0.70 mmol/L) than in the control (2.93 ± 0.39 mmol/L). By contrast, the level of serum insulin was lower in group B (5.641 ± 0.573 μIU/mL) than in the control (7.578 ± 0.539 μIU/mL). Histological studies by hematoxylin and eosin (H&E) revealed a loss of β-cells in the pancreas from pigs treated with 170 mg/kg alloxan. Immunolocalization studies showed a decrease in insulin reactivity in this treatment group as well. To conclude, our model holds promise in future studies of diabetes drug testing and islet xenotransplantation.
authors.affiliation: a Department of Animal Science and Animal Medical , College of Agriculture, Yanbian University , Yanji , China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Guo
authors.forename: Hai-Ying
authors.initials: HY
authors.lastname: Zhu
authors.forename: Long
authors.initials: L
authors.lastname: Jin
authors.forename: Qing-Shan
authors.initials: QS
authors.lastname: Gao
authors.forename: Jin-Dan
authors.initials: JD
authors.lastname: Kang
authors.forename: Cheng-Du
authors.initials: CD
authors.lastname: Cui
authors.forename: Xi-Jun
authors.initials: XJ
authors.lastname: Yin
mesh_terms: D000496:Alloxan; D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D003001:Cloning, Molecular / Q000379:methods*; D003921:Diabetes Mellitus, Experimental; D004195:Disease Models, Animal; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D007328:Insulin / Q000097:blood; D007515:Islets of Langerhans / Q000737:chemistry; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics*; D018675:Toxicity Tests
chemical_list: D001786:Blood Glucose; D007328:Insulin; D000496:Alloxan
keywords: Alloxan; Cloned; Diabetes model; Miniature pig