• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.65-70
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• 2004

In mammals, male and female germline stem cells are derived from primodial germ cells. Despite many efforts to identify stem cells from gonads, there has been little successe to identify germline stem cells yet. In this study, we isolate and characterized porcine germline stem cells using only stem cell markers that are prevalently expressed in various tissues. Gonadal cells derived from both male and female formed colonies and showed AP activities and different lectin binding properties. Pluripotency of germline stem cells was also identified by positive signals against putative stem cells markers such as SSEA-1 and SSEA-3. In addition, nestin was also found in primary gonad cells that have a similar morphology to the AP-positive cells. The nestin expression suggests that the germline stem cells may have similar expression of the prevalent stem cell markers found in other tissues. The demonstration of nestin expression together with pluripotent cell markers calls further investigation of the possible differentiation of nestin-positive cells into neurons.

• Reproductive and Developmental Biology
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• v.33 no.3
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• pp.119-123
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• 2009

The two dimensional quantitative structure-activity relationships (2D-QSARs) models concerning the binding affinity constants ($p[Od.]_{50}$) between 2-cyclohexyltetrahydropyrane and 2-cyclohexyltetrahydrofurane analogues as substrates, and bovine odorant binding protein (bOBP) as receptor were derived by multiple regression analyses method and discussed. The statistical quality of the optimized 2D-QSAR model (5) was good (r=0.907). From the model, the binding affinity constants ($p[Od.]_{50}$) were dependent upon the optimal value ($(TL)_{opt.}$=2.737) of total lipole (TL) of substrate molecules. Based on these findings, the high active compounds predicted by optimized 2D-QSAR model (5) were 2-(dimethylcyclohexyl)tetrahydropyrane molecule and their isomer molecules. The binding affinity constants regarding bOBP of the tetrahydrofuryl-2-yl family compounds were dependent upon the hydrophobicity (logP) of whole substrate molecules. In any case of porcine odorant-binding proteins (pOBP), the constants were dependent upon the hydrophobicity (${\pi}x={\log}P_X-{\log}P_H$) of substituents (R) in substrate molecules. Also, from the optimal values of hydrophobic constant, the hydrophobicity for bOBP influenced ca. twice time bigger (bOBP>pOBP) than that for pOBP.

• Journal of Veterinary Clinics
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• v.26 no.1
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• pp.29-35
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• 2009

This study was to determine the effects of ascorbic acid and alpha-tocopherol on the attenuation of an ischemia-reperfusion injury (IRI) after renal auto-transplantation in a pig model. In the treatment group, three pigs were subjected to a renal auto-transplantation followed by the administration of ascorbic acid and alpha-tocopherol and the flushing of ascorbic acid plus hepa-saline solution. Otherwise, the control group used only flushing of hepa-saline solution. Blood samples were collected from these pigs for measurement of serum blood urea nitrogen (BUN) and creatinine values on the day before surgery and day 1, 3, 5 and 7 after surgery. The kidneys were taken for histopathological evaluation following euthanasia on day 14 after surgery. Serum creatinine and BUN values showed a significantly difference between control and treatment group on day 1, 3 and 5 (P<0.05). In histopathologic findings, treatment group showed less damage than that of the control group on the basis of renal tubular damage. As a result, this study suggests that the exogenous ascorbic acid and alpha-tocopherol pretreatment therapy with ascorbic acid irrigation-aspiration has a role of attenuation of renal I/R injury and recovery of renal function in a pig transplantation model.

• Journal of Life Science
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• v.28 no.4
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• pp.389-397
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• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.179-184
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• 2007

Culture method of somatic cells is one of the important factors in the production of transgenic pigs by somatic cell nuclear transfer. In this study, we established an efficient culture method of porcine fetal fibroblasts. Porcine fetal fibroblasts were isolated from 33-day-old fetuses. The proliferation of porcine fetal fibroblasts was analyzed by different serum types and culture media. The cultures in medium supplied 15% ES screened FBS showed faster increase in cell number than 15% FBS. Also, fetal fibroblasts have been propagated continuously for $7{\sim}8$ passages in ES modified DMEM and DMEM medium. We transfected $PGK-neo^r$ vector (pKJ2) into porcine fetal fibroblasts to estimate colony formation in this culture condition. The formation of colonies was confirmed in the medium containing $300\;{\mu}g/ml$ G418 at 12 day. These data show that this culture system can be used screening of porcine somatic cells transfected transgene.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.563-567
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• 2009

This study compared the instrument performance and tissue healing of a steel scalpel with a $CO_2$ laser in an animal uterine surgery model. Five Landrace and Yorkshire mixed breed pigs were used. Two symmetrical incisions were made in the uterine of each pig. One incision was made on the left side of the uterine horn using a steel scalpel, while the other incision was performed on the right side using a $CO_2$ laser with an 8W output power. Each instrument was evaluated clinically for speed, ease of incision, and extent of bleeding. An ovariohysterectomy was performed at 21 days after the surgical procedure for a histological examination. The scalpel was an easier instrument to use in the confines of the uterine tissue, compared with the laser. However, there is no significant difference between the two groups. The amount of bleeding was less in the laser group but the time of the incisions was shorter with the scalpel. Postoperative uterus adhesion in the $CO_2$ laser incisions was lower than the scalpel incisions. Scalpel incisions showed complete restoration of the epithelium and endometrial gland. On the other hand, the laser incisions showed incomplete restoration of the epithelium and endometrial gland. Although the scalpel produced less damage to the uterine tissue and was easier to handle than the $CO_2$ laser, it did not provide hemostasis that was helpful for use on highly vascular tissue. The $CO_2$ laser provided good hemostasis but delayed wound healing.

• Korean Journal of Veterinary Research
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• v.43 no.2
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• pp.301-305
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• 2003

To evaluate the effects of Zoletil (tiletamine- zolazepam) that have been widely used for the chemical restraint and anesthesia of primates, on physiologic alteration, blood gas analysis and anesthetic duration in the Japanese macaque(Macaca fuscata), this study was carried out. Zoletil was administered by intramuscular injection. Evaluation of temperature, heart rate, respiration rate and blood gas analysis were performed before administration and at 1, 10, 20, 30, 40, 50 and 60 min after administration, and induction and maintenance time was recorded. There was no significant difference in heart rate, $PCO_2$, $PO_2$ after Zoletil administration rut temperature, respiraticn rate, pH were significant difference compared with these of Mere administration. The induction time was $2.5{\pm}1.0min$ and maintenance time was $86{\pm}23.2min$. It was considered that Zoletil could be usefully used for the sedation and immobilization of Japanese macaque reared in zoological garden.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.53-59
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• 2003

For somatic cell nuclear transfer in Hanwoo, fetal fibroblast cell lines were established from 35, 50, 70 and 90-day fetuses of Korean native cattle. The sex of these fetal fibroblast cells were analyzed by PCR using Y-specific primers and confirmed that two cell lines were female and the other two cell lines were male. Karyotyping of these cell lines indicates that the chromosome numbers of fetal fibroblast cells were not affected by passage number and more than 80% of fetal fibroblast cells have normal chromosome number. To evaluate Go stage in cell cycle of fetal fibroblast cells, Western blotting was performed to detect the expression level of PCNA which is known to be expressed in all cell cycle stages except G$_{0}$ stage. Following serum starvation or confluent culture for 7 days, fetal fibroblast cells were effectively reached to G$_{0}$ stage. The cell cycle was resumed after culture of these Go stage-fetal fibroblast cells with normal medium. These results indicates that fetal fibroblast cells originated from Hanwoo were successfully isolated and culture system and induction of cell cycle of these cells were established for somatic cell nuclear transfer in Hanwoo.woo.

• Journal of Animal Science and Technology
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• v.46 no.1
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• pp.77-82
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• 2004

This study was carried out to establish the purification protocol of angiogenin(ANG) from bovine milk. The purification of ANG from bovine milk was performed by using cation chromatography, high-performance liquid chromatography and gel-filtration. We obtained the ANG protein have the molecular weight of about 14 kD by SDS-PAGE analysis. This protein was confirmed as ANG by Nlb-terminal sequence analysis of the first 15 amino acids. Identified amino acids revealed the protein to be identical to that previously reported for bovine ANG.

• Journal of Veterinary Clinics
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• v.23 no.3
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• pp.279-285
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• 2006

Thirty dairy cows with pododermatitis were selected and treatment effect of ozone ointment for bovine pododermatitis was investigated. In addition, bactericidal effect of ozone ointment on etiological agent of bovine pododermatitis was evaluated. The pathohistological examination for the pododermatitis, according to application with ozone ointment was investigated. Thirty dairy cows were divided two groups: control group(vaseline group: 15 cows), treatment group(ozone ointment group: 15 cows). Various parameters were evaluated in terms of the lameness score, swelling score, lesion score, WBC, neutrophil, pathohistological finding, and antimicrobial action. As compared with vaseline group, ozone ointment group revealed significant decrease of lameness(p<0.05), swelling(p<0.01) and lesion score(p<0.05) were shown in hoof lesions on 14 days after application. In hematological findings, WBC count revealed slightly high values within normal range before treatment, however, this was 1Corresponding author improved on 14 days after application of ozone ointment. The number of neutrophils was slightly higher than that of normal, however, this was improved on 14 days after application of ozone ointment. In pathohistological findings, normal dermal tissue was found in tissues with pododermatitis on 14 days after application of ozone ointment. In antimicobial action, marked decrease rate of bacteria was observed in feet of all cases treated with ozone ointment. The decreasing rate of bacteria in anaerobic culture was higher than that in aerobic culture.