Establishment and identification of cell lines from type O blood Korean native pigs and their efficiency in supporting embryonic development via somatic cell nuclear transfer

Due to their similarities with humans in anatomy, physiology, and genetics miniature pigs are becoming an attractive model for biomedical research. We aim to establish and evaluate blood type O cells derived from Korean native pig (KNP), a typical miniature pig breed in Korea. Ten cell lines derived from 8 KNP piglets and one adult female KNP (kidney and ear tissues) were established. To confirm the presence of blood type O, genomic DNA, fucosyltransferase (FUT) expression, and immunofluorescence staining were examined. Additionally, fluorescence-activated cell sorting and somatic cell nuclear transfer were performed to investigate the normality of the cell lines and to evaluate their effectiveness in embryo development. We found no significant bands corresponding to specific blood group A, and no increase in FUT expression in cell lines derived from piglets No. 1, No. 4, No. 5, No. 8, and the adult female KNP; moreover, they showed normal levels of expression of α 1,3-galactosyltransferase and cytidine monophosphate-N-acetylneuraminic acid hydroxylase. There was no significant difference in embryo development between skin and kidney fibroblasts derived from the blood type O KNPs. In conclusion, we successfully established blood type O KNP cell lines, which may serve as a useful model in xenotransplantation research.

Disagreement between direct and indirect blood pressure measurements obtained from minipigs / 대한수의학회지

Blood pressure (BP) measurement plays a pivotal role in veterinary medicine for diagnosing cardiovascular disorders and monitoring anesthesia of animals. Although indirect BP measurement has been widely applied to monitor BP because of its convenience and non-invasiveness, it is still unclear whether indirect BP measurement is compatible with direct BP measurement in minipigs. In addition, the effect of animal posture during BP measurement is not well understood in minipigs despite its importance to cardiovascular performance. Therefore, both systolic and diastolic arterial BPs in minipigs were measured via femoral artery catheterization for direct BP measurement and using a compressive cuff as an indirect BP measurement under the dorsal or right lateral recumbent postures. Numerical values were processed by the Bland-Altman method to calculate the bias ± SD and the limits of agreement (LOA). In accordance with the American College of Veterinary Internal Medicine guidelines, the results between direct and indirect BP measurements were determined as apparent disagreements in both systolic and diastolic arterial BPs under all postures because of large bias ± SD and wide LOA. The results of the present will help prevent misinterpretation of the anesthetized patient's condition during monitoring of BP by indirect measurement.

Effects of Salvia miltiorrhiza-Volatile Oil of Dalbergia odorifera on Blood Lipid and Blood Coagulation System in Coronary Heart Disease Myocardial Ischemia Miniature Swine with Blood Stasis Syndrome / 中国药房

OBJECTIVE:To observe the effects of Salvia miltiorrhiza-volatile oil of Dalbergia odorifera on blood lipid and blood coagulation system in coronary heart disease myocardial ischemia miniature swine with blood stasis syndrome. METHODS:A total of 18 swine were randomly divided into sham operation group (routine feed),model group (routine feed) and S.miltiorrhiza-volatile oil of D. odorifera group(1 g/kg S. miltiorrhiza+0.1 mL/kg oil of D. odorifera,mixing administration),for consecutive 8 weeks. At 4th week,coronary heart disease myocardial ischemia model of blood stasis syndrome was established by Ameriod coarctation ring implantation in other 2 groups except for sham operation group. At 8th week,the syndrome of the model was observed according to coronary angiography and the macroscopic indications. At 2th,6th,8th week,intravenous blood was collected to test the serum levels of TC,TG,LDL-C,HDL-C,apolipoprotein A1(apoA1),apoB,PT,APTT,TT and FIB in miniature swine. RESULTS:At 6th,8th week,compared with sham operation group,serum levels of TC,TG,LDL-C,apoB and FIB were increased in model group,while PT,APTT and TT were shortened and the levels of HDL-C and apoA1 were decreased,with statistical significance(P<0.05 or P<0.01). Compared with model group,the serum levels of TC,TG,LDL-C and apoB were decreased in S. miltiorrhiza-volatile oil of D. odorifera group,while PT,APTT and TT were prolonged and the levels of HDL-C and apoA1 were increased, with statistical significance (P<0.05 or P<0.01). CONCLUSIONS: The S.miltiorrhiza-volatile oil of D. odorifera could reduce blood lipid and improve blood coagulation system index disorder in coronary heart disease myocardial ischemia miniature swine with blood stasis syndrome.

Relationship between prescription and syndrome on mini-swine phlegm-stasis syndrome of coronary heart disease / 药学学报

This study was conducted to investigate the effects of Danlou (correspondence between prescription and syndrome) tablet and Shengmai capsule (non-correspondence between prescription and syndrome) on mini-swine phlegm-stasis syndrome of coronary heart disease (CHD). 24 mini-swines were randomly divided into normal control group, model group, Danlou tablet group (0.24 g·kg-1) and Shengmai capsule group (0.14 g·kg-1). Phlegm-stasis syndrome of coronary heart disease was established by high-fat feeding and coronary intervention balloon injury. After 8 weeks of administration, blood lipid levels and blood rheology was detected. Echocardiography was used to examine the changes in heart function, and the extent of infarction was determined by nitro blue tetrazolium (NBT) staining method. The main symptoms, accompanied symptoms, tongue and pulse signs of the coronary heart disease mini-swine with phlegm-stasis syndrome were observed according to the symptom-graded scoring method. The results showed that Danlou tablet decreased serum total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) (P P P P P P P P P P P P < 0.05). However, Shengmai capsule failed to show therapeutic effects on blood lipid metabolism, the myocardial infarction area and primary symptom and syndrome score. The results suggest that as a drug for the treatment of Qi and Yin deficiency syndrome of CHD, Danlou tablet has limited therapeutic effects on phlegm-stasis syndrome of CHD. Only by the prescription correspondence with syndrome, using drug for the treatment of phlegm-stasis syndrome of CHD to treat phlegm-stasis syndrome of CHD, the prescription has a comprehensive therapeutic effect.

Brazilian minipig as a large-animal model for basic research and stem cell-based tissue engineering. Characterization and in vitro differentiation of bone marrow-derived mesenchymal stem cells

Stem cell-based regenerative medicine is one of the most intensively researched medical issues. Pre-clinical studies in a large-animal model, especially in swine or miniature pigs, are highly relevant to human applications. Mesenchymal stem cells (MSCs) have been isolated and expanded from different sources. Objective: This study aimed at isolating and characterizing, for the first time, bone marrow-derived MSCs (BM-MSCs) from a Brazilian minipig (BR1). Also, this aimed to validate a new large-animal model for stem cell-based tissue engineering. Material and Methods: Bone marrow (BM) was aspirated from the posterior iliac crest of twelve adult male BR1 under general anesthesia. MSCs were selected by plastic-adherence as originally described by Friedenstein. Cell morphology, surface marker expression, and cellular differentiation were examined. The immunophenotypic profile was determined by flow cytometry. The differentiation potential was assessed by cytological staining and by RT-PCR. Results: MSCs were present in all minipig BM samples. These cells showed fibroblastic morphology and were positive for the surface markers CD90 (88.6%), CD29 (89.8%), CD44 (86.9%) and negative for CD34 (1.61%), CD45 (1.83%), CD14 (1.77%) and MHC-II (2.69%). MSCs were differentiated into adipocytes, osteoblasts, and chondroblasts as demonstrated by the presence of lipidic-rich vacuoles, the mineralized extracellular matrix, and the great presence of glycosaminoglycans, respectively. The higher gene expression of adipocyte fatty-acid binding protein (AP2), alkaline phosphatase (ALP) and collagen type 2 (COLII) also confirmed the trilineage differentiation (p<0.001, p<0.001, p=0.031; respectively). Conclusions: The isolation, cultivation, and differentiation of BM-MSCs from BR1 makes this animal eligible as a useful large-animal model for stem cell-based studies in Brazil. .

Periodontal ligament cells from miniature swine grow on a hydroxyapatite scaffold / 中国组织工程研究

BACKGROUND:Periodontal tissue engineering technology provides new ideas and new ways for periodontitis-induced bone defect repair. OBJECTIVE:To develop a culture model for the periodontal ligament cells of miniature swine, which was constructed with hydroxyapatite, to investigate the biocompatibility with hydroxyapatite. METHODS:Periodontal ligament cells from miniature swine were harvested by using tissue explant method. Immunofluorescence was used to detect the expression of stromal cel antigen 1 in the periodontal ligament cells of miniature swine. The third passage cells were co-cultured with a three-dimension hydroxyapatite scaffold, and the biological characteristics of the cells were observed under a scanning electron microscope at days 1, 3, 7 of co-culture. RESULTS AND CONCLUSION:The pirmary miniature swine periodontal ligament cells grew wel , and they were positive for stromal cel antigen 1. Under the scanning electron microscope, the periodontal ligament cells of miniature swine grew wel on the hydroxyapatite scaffold at days 1, 3, 7 of co-culture. These prove that the miniature swine periodontal ligament cells, which can be separated using tissue explant method and cultured successful y in vitro, can grow wel on the hydroxyapatite scaffold.

Experimental study of the protective effect of PTX on bile duct in grafts against IR injury in the animal model of OLTx in miniature swines / 第三军医大学学报

Objective To investigate the protective effect of pentoxifylline (PTX) on bile duct in grafts against ischemia-reperfusion (IR) injury and its potential mechanisms in the animal model of OLTx in miniature swines. Methods After PTX was added to the preservation solution and administrated through hepatic artery, changes of blood flow, activity of ATPase, apoptosis of epithelial cells, and pathomorphology of the bile duct were investigated in the animal model of OLTx in miniature swines. The animals were divided into IR, IR+NS, and IR + PTX groups. Results The results showed that the incidence of biliary necrosis was significantly lower in IR+PTX group than that in IR and IR +NS groups. When compared with those in IR and IR +NS groups, GGT and ALP levels, numbers of apoptosis cells, and scores of pathomorphology were significantly decreased in IR+PTX group, while blood flow, activity of Na +-K +ATPase, Ca 2+ -ATPase of bile duct in grafts were significantly elevated. Conclusion These results suggest that PTX has multiple antagonistic effects against IR injuries of the bile duct. The effects of PTX on bile duct of grafts against IR injuries are possibly correlated with the improvement of the microcirculation, preservation of the ATPase activity of the bile duct, and suppression of the apoptosis of the bile duct cells.

Effects of Shuangshen Ningxin capsule on ventricular remodeling and wall motion in myocardial ischemic miniature swine induced by thrombus / 中国药理学通报

Aim To observe the improved effects of Shuangshen Ningxin capsule on ventricular remodeling and wall motion in myocardial ischemic miniature swine induced by thrombus.Methods Myocardial ischemic model miniature swine induced by self-thrombus via cardiac catheter in left anterior descending coronary artery(LAD)were administrated Shuangshen Ningxin capsule for 6 days.The changes of cardiac form,left heart systolic and diastolic function,left ventricular wall motion were observed by the technology of conventional ultrasound and Doppler tissue imaging.Results 6 days after myocardial ischemia and administration,miniature swine of model group showed ventricular wall thinning,chambers heart dilating,ventricle remodeling and wall motion weak,but Shuangshen Ningxin capsule obviously lowered left ventricular internal diameter at end-diastole(LVIDd),left ventricular internal diameter at end-systole(LVIDs),end-systolic volume(ESV),end-diastolic volume(EDV)and isovolumic relaxation time(IVRT),and increased ejection fraction(EF),tissue velocity and tissue trace of left ventricle anterior wall cardiac apex segment.Conclusion Shuangshen Ningxin capsule has anti-myocardial ischemia effect by improving left ventricular remodeling,increasing left cardiac work,improving left heart systolic and diastolic function and increasing left ventricular wall motion.