Adipose tissue-derived mesenchymal stem cells (ASCs) are very interesting in several laboratory animals and humans because they are easy to harvest and expand to generate millions of cells from a small quantity of fat. ASCs are known as useful materials for clinical applications in human cell therapy and as a donor cell in somatic cell nuclear transfer (SCNT). Here, we investigated if 1) minipig ASCs can be isolated, self-renewed and differentiated into multiple tissue lineages, 2) ASCs can be a suitable donor cell type for generation of cloned pig. In order to isolate ASC, adipose tissues were collected from inguinal region of a 6-year-old female minipig. The ASCs were attached to the culture dish with a fibroblast-like morphology. They expressed cell-surface marker characteristics of stem cell, underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. To investigate its potential as donor cell for cloning, we respectively carried out SCNT using ASC, adult skin fibroblast (ASF) and fetal fibroblast (FF) derived from same minipig. The ratio of blastocysts to 2-cell embryos and total cell number of blastocysts were monitored as experimental parameters. In results, cleavage and developmental competence to blastocysts rate showed no significant difference among the three groups. On the other hand, total cell numbers of blastocysts derived from ASC and FF were significantly higher than in ASF (89±7.9 and 105±5.5 vs. 57.5±5.2, respectively). Our results demonstrated that ASC have potential compared to ASF and FF in terms of the in vitro development and blastocyst formation ability. In further study, we will investigate the in vivo developmental ability of ASC as donor cell for pig cloning. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program, TS Corporation and Optifarm Solution.

Minipigs are regarded as one of the most important laboratory animal in that anatomical and physiological properties are similar to human and their reproduction efficiency is relatively higher compared to other large animal species. Particularly, several diseases that cannot be mimicked in rodent models are successfully occurred or induced in pig models therefore it has been interested in a valuable model for human diseases. Pigs are also ‘standard’ species in xenotransplantation research. To maximize experimental outcome using minipigs, establishment and management of proper animal facility, right animal husbandry and control of pathogens are very important. In this review, we summarized several international guidelines related with minipigs published by several companies or governments and discuss optimal conditions for providing informative ideas to the researchers who want to use minipigs in their future studies.

Due to the similarities of their anatomical morphology and physiology to those of humans, pigs have increasingly become useful models for toxicological and pharmacological studies. This study was conducted in order to evaluate the quality control level through hematological features of minipigs. Thus, we attempted to determine whether there are differences in hematological parameters between male and female minipigs. No significant differences in the levels of erythrocytes, hematocrit, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), leukocytes (WBC), and serum electrolytes were observed between male and female minipigs. However, leukocytes were higher than reference range in both sexes (26.46±2.06 vs 11.6±5.4 for male; 23.58±3.36 vs 10.0±2.0 for female). In differential leukocyte counts, no significant differences in leukocyte parameters were observed between sexes. However, neutrophils were higher than reference range in both sexes (60.65±13.92 vs 33.1±18.9 for male; 46.84±12.80 vs 15.4±5.6 for female). In serum biochemical analysis, no significant differences in biochemical parameters were observed between male and female minipigs. However, aspartate aminotransferase (AST) and alkaline phosphatase (ALT) were higher than reference range, while alanine aminotransferase (ALP) was lower than reference range. In urine analysis, pH of male and female was 8.3±1.0 and 6.4±1.6, showing a higher value in male minipigs. These data indicate that quality control of minipigs must take precedence in conduct of toxicological and pharmacological studies.